Novel plant receptor-like kinases

ABSTRACT

The present invention is related to plant molecular biology. Particularly it is related to nucleic acids and methods for conferring disease resistance in plants. It is also related to potato receptor-like kinases (PRKs) and PRK cDNA or nucleic acid sequences and their gene products for conferring enhanced resistance to pathogens and pests. The invention is further related to novel receptors and ligands and their use in detecting plant-pathogen interactions.

TECHNICAL FIELD OF THE INVENTION

[0001] The present invention is related to plant molecular biology. Particularly it is related to nucleic acids and methods for conferring disease resistance in plants. It is also related to potato receptor-like kinases (PRKs) and PRK cDNA or nucleic acid sequences and their gene products as well as fragments and derivatives thereof useful in conferring enhanced resistance to pathogens and pests. The invention is further related to novel receptors and ligands and their use in detecting plant-pathogen interactions.

BACKGROUND OF THE INVENTION

[0002] Potato is the 4^(th) major food crop of the world and expanding. Potato's susceptibility to pests and diseases makes the crop the number two user of agricultural pesticides worldwide, following cotton. Erwinia carotovora is the etiological agent of soft rot disease and can attack a wide range of economically important crops including potato (Pérombelon and Kelman, 1980). The production of extracellular plant cell wall-degrading enzymes, including cellulases, pectinases and proteases is central to virulence of E. carotovora. These enzymes both produce the maceration symptoms in infected plant tissues and release nutrients for bacterial growth (Collmer and Keen, 1986; Kotoujansky 1987; Pirhonen et al., 1991). Many of the plant cell wall-degrading enzymes have been shown to trigger plant defense responses, probably by releasing cell wall fragments active as elicitors (Davis and Ausubel, 1989; Davis et al., 1984; Palva et al., 1993; Vidal et al., 1997, 1998). It has been previously demonstrated that cell-free culture filtrates (CF) containing the cell wall-degrading enzymes of E. carotovora subsp. carotovora, as well as preparations containing single enzymes, induce several pathogenesis-related genes in plants (Norman et al., 1999, Norman-Setterblad et al., 2000; Palva et al., 1993; Vidal et al., 1997, 1998). In addition, it has been shown that several of these defense-related genes are also responsive to oligogalacturonides (Norman et al., 1999).

[0003] Plant receptor-like kinases (RLKs) are proteins with a predicted signal sequence, single transmembrane region, and cytoplasmic kinase domain. Plant RLKs show serine/threonine kinase specificity. Based on the structure of the putative extracellular domains, plant receptor-like kinases (RLKs) have been classified into several major classes (Braun and Walker, 1996; Walker, 1994). These include (i) the S-domain RLKs which contain extracellular domains homologous to the S-locus glycoproteins of Brassicaceae (Nasrallah et al., 1993; Walker and Zhang, 1990, Stein et al., 1991), (ii) the leucine rich repeat (LRR) RLKs such as Xa21 from rice, TMK1 and RLK5 from Arabidopsis (Chang et al., 1992; Song et al., 1995; Walker, 1993) and (iii) RLKs with the epidermal growth factor-like repeat (EGF) such as pro25 and the WAKs from Arabidopsis (He et al., 1999; Kohorn et al., 1992). Moreover, several RLKs with different types of extracellular domains have been identified recently (reviewed by Satterlee and Sussman, 1998).

[0004] The expression of plant RLK genes have shown diverse patterns, while some of them have displayed expression only in vegetative tissues (Kohorn et al., 1992), others were expressed only in reproductive tissues (Goring et al., 1992; Stein et al., 1991) and some have been shown in both vegetative and reproductive tissues (Pastuglia et al., 1997). Some of the RLK genes are responsive to pathogens and elicitors, including PvRK20-1 from Phaseolus vulgaris (Lange et al., 1999), SFR2 from Brassica olearacea (Pastuglia et al., 1997), Wak1 (He et al., 1998) and RLKs (Du and Chen, 2000) from Arabidopsis thaliana and the disease resistance gene Xa21 from rice (Song et al., 1995).

[0005] In plants very little is known about the nature of the ligands interacting with serine-threonine RLKs. Recently, it has been shown that the extracellular domain of a RLK from Arabidopsis, BRI1, perceives brassinosteroids (He et al., 2000). On the other hand, in animals it has been shown that the interleukin 2 and the epidermal growth factor receptors are up regulated by their own ligands (Clark et al., 1985; Deeper et al., 1985).

[0006] In order to fully understand the mechanisms of plant disease resistance and provide plants with enhanced disease resistance to pathogens and even herbivores (insects pests) novel means for studying the interaction between ligand and receptor in plant-pathogen interaction are needed.

[0007] The present invention provides a solution to said problem. Novel potato receptor-like kinases (PRKs) as well as a new expression pattern are described: PRK is induced by the potato pathogen Erwinia carotovora as well as short oligouronides, which may be the elicitors released by Erwinia. Oligouronides may constitute the ligands for the novel receptor. Oligouronide receptor has not been described previously. The structural identity of PRKs and their induction pattern suggested that they constitute part of the early response of potato E. carotovora infection.

[0008] An embodiment of the present invention is to provide isolated nucleic acid sequences comprising polynucleotides encoding receptor-like protein kinases, which comprise preferably in the extracellular domain one or more cystein repeats, characterized in that potato receptor-like kinase (PRK)-like nucleic acid sequences are capable of encoding potato receptor-like kinases (PRKs) substantially homologous to gene products of PRK (SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4) or fragments or derivatives thereof and having substantially the same properties or functions as SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4.

[0009] An embodiment of the present invention is to provide new methods for conferring resistance to pathogens and herbivores (insect pests) producing/releasing elicitors of the present invention.

[0010] Another embodiment of the present invention is to provide new means and methods for making it possible to initiate the defense mechanisms during the early stages of the plant-pathogen interaction.

[0011] Another embodiment of the present invention is to provide means carrying out said methods. The means are potato receptor-like kinase (PRK) cDNA and PRK nucleic acid sequences, fragments and derivatives thereof as well as their complementary strands and PRK gene products expressed by the PRK nucleic acid sequences of the present invention.

[0012] Another embodiment of the present invention is to use PRK gene products, fragments and derivatives thereof for technically modifiying the expression of a gene or a modified gene/a natural variant of the gene to sensitize the plant to pathogen perception and to generate enhanced resistance.

[0013] Another embodiment of the present invention is to use the ligand of the receptor of the present invention for spraying, inoculating, spreading, applying or by other means the crop plants means to enhance disease resistance.

[0014] Another embodiment is to use PRK-like nucleic acid sequences as well as their expression products for manufacturing transgenic plant cells or plants with enhanced pathogen resistance.

[0015] Another embodiment is to use PRK-like nucleic acid sequences and PRK-like gene products and derivatives and fragments thereof as growth regulators, for induction of development, for detecting different stress conditions. Different stress conditions can be caused e.g. by pathogens and pests, mechanical, chemical or physical stress.

SUMMARY OF THE INVENTION

[0016] The present invention is related to related to potato receptor-like kinases (PRKs) and PRK-like nucleic acid sequences and fragments and derivatives thereof and their products useful in conferring enhanced resistance to pathogens and pests.

[0017] The present invention provides four isolated nucleic acid sequences comprising SEQ ID:5, SEQ ID:6, SEQ ID:7, SEQ ID:8.

[0018] The said nucleic acid sequences comprise genes encoding novel potato receptor-like kinases (PRKs) (PRK-1, PRK-2, PRK-3 and PRK-4), SEQ ID:1, SEQ ID:2, SEQ ID:3, SEQ ID:4. They are proposed to belong to a new class of receptor-like protein kinases. Potato PRKs are 41-55% i.e. highly distinct in the extracellular domain from other receptor-like protein kinases of the same class, PvRK20-1 from Phaseolus vulgaris (Lange et al., 1999) and genes from the genome project of Arabidopsis thaliana (Accession numbers CAB38617, CAB81062, CAA18704).

[0019] A new expression pattern is described: PRK is induced by the potato pathogen Erwinia carotovora as well as short oligouronides, which may be the elicitors released by Erwinia. Oligouronides may constitute the ligands for the receptor. Oligouronide receptor has not been described previously.

[0020] The present invention is related to isolated nucleic acid sequences comprising polynucleotides encoding receptor-like protein kinases, which comprise preferably in the extracellular domain one or more cystein repeats, characterized in that potato receptor-like kinase (PRK)-like nucleic acid sequences are capable of encoding potato receptor-like kinases (PRKs) substantially homologous to gene products of PRK (SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4) or fragments or derivatives thereof and having substantially the same properties or functions as SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4. The potato receptor-like kinase (PRK)-like nucleic acid sequences are capable of hybridizing with SEQ ID:5, SEQ ID:6, SEQ ID:7 or SEQ ID:8 or complementary strands thereof under defined conditions.

[0021] The nucleic acid sequences comprise SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4 obtainable from potato under defined conditions. The extracellular domains comprise a conserved bi-modular pattern of one or more cysteine repeats. The expression of genes encoding for gene products SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4 is induced by Erwinia carotovora.

[0022] The expression of genes encoding for gene products SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4 is induced by oligouronides. Potato receptor-like kinases (PRKs) function as receptors for ligands released during plant stress conditions by pathogens. Potato receptor-like kinases (PRKs) are formed by alternative splicing. Potato receptor-like kinases (PRKs) are involved in signal perception during potato defense responses against Erwinia carotovora. The expression of potato receptor-like kinases (PRKs) is incuced by response to elicitors, which can be oligouronides. Or oligogalacturonides.

[0023] The PRK-like gene products are polypeptides comprising in their extracellular domains a conserved bi-modular patterns of one or more cysteine repeats. The potato receptor-like kinase (PRK)-like gene products comprise polypeptides having amino acid sequences substantially homologous with SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4. The potato receptor-like kinase (PRK)-like gene products are polypeptides substantially similar to the gene products encoded by SEQ ID:5, SEQ ID:6, SEQ ID:7 or SEQ ID:8.

[0024] The present invention is related to the method for preventing plant diseases or enhancing disease resistance to pathogens or herbivores by transforming a plant with a DNA construct comprising nucleic acid sequences encoding potato-receptor like kinase (PRK)-like gene products or fragments thereof functionally combined with regulatory sequences. The present invention is also related to DNA constructs, expression vectors and host cells comprising the DNA sequences of the present invention.

[0025] The present invention is related to a method for conferring resistance to pathogens in a plant, preferably potato, so that the method comprises introducing into the plant a recombinant expression construct comprising a plant promoter operably linked to a potato receptor-like kinase (PRK)-like nucleotide sequence or derivatives or fragments thereof encoding a potato receptor-like kinase (PRK)-like gene product.

[0026] The present invention is as defined in the claims of the present invention.

SHORT DESCRIPTION OF THE FIGURES

[0027]FIG. 1 depicts the comparison of the deduced amino acid sequences of PRKs. Identical amino acids are highlighted with black and similar amino acids with gray. Dashes indicate gaps introduced to improve the alignment. The two hydrophobic regions flanking the putative extracellular domain are double underlined and asterisks indicate the region of basic residues. The numbered brackets indicate putative glycosylation sites. Note that under the brackets number 4 and 6 the putative glycosylation sites are missing from PRK-3 and PRK-1, respectively. The cDNA clones corresponding to the PRKs were sequenced at the DNA Synthesis and Sequencing Unit of the Institute of Biotechnology, Helsinki, Finland, using ABI 377 system. The alignment was performed using PILE UP from the Genetics Computer Group (GCG) software package.

[0028]FIG. 2 depicts the structural analysis of PRKs. (a) Comparison of the kinase domains of PRK-1, 2, 3 and 4 with the kinase domain of SFR2 (Pastuglia et al., 1997; accession number P93068), IRK1 (Kowyama et al., 1996; accession number Q40096), PvRK20-1 (Lange et al., 1999; accession number AF078082) and a putative Arabidopsis thaliana (At) receptor-like kinase (Bevan et al., Unpublished data; accession number 065470). The 11 characteristic subdomains of kinases are indicated by roman numbers, and the 15 invariant amino acids are indicated by asterisks and highlighted with black. The two regions in subdomains VI and VIII indicative of serine-threonine kinases are boxed and shaded in gray. Consensus indicates the conserved residues in all sequences shown. (b) Alignment of the extracellular domains of PRK-2, 4 and 3 at the region where PRK-3 lacks 25 amino acids. The cDNA and the translated amino acid sequences near the possible splice sites are shown and the 25 amino acids present in PRK-2 and 4 but not in 3 are highlighted in black. Conserved sequences of splice sites (Brown and Simpson, 1998) are highlighted in gray. An asterisk indicates a conserved cysteine and the bracket indicates a putative N-glycosylation site. The nucleotides generating a different codon in PRK-3 are underlined, as is the amino acid that is altered as a result of the splicing event. (c) Alignment of the extracellular domains of PRK-1, 2, 3 and 4, PvRK20-1 and three genes from the Arabidopsis genome project (Bevan et al., Unpublished data) here named as At1 (accession number CAB38617), At2 (accession number CAB81062) and At3 (accession number CAA18704). Cysteine amino acids are highlighted with black and the numbers above the double line indicate the number of amino acids between two cysteine residues. Putative glycosylation sites are highlighted with gray. Consensus indicates the conserved residues in all sequences shown.

[0029]FIG. 3 depicts Southern blot analysis of PRKs. Genomic DNA samples (5 μg) from S. tuberosum digested with the indicated restriction enzymes were separated by electrophoresis in a 0.8% agarose gel. Hybridization and washes were done according to Sambrook and Russell (2001). A fragment of the first 450 bases corresponding to the 5′ end of PRK-2 cDNA was used as probe labeled with [α-³²P]dCTP by random priming (Amersham International, UK). λ DNA digested with PstI together with a 100-basepair ruler were used as molecular markers.

[0030]FIG. 4 depicts the accumulation of PRK mRNAs in potato tissues in response to E. carotovora culture filtrate. (a) Accumulation of PRK mRNAs in leaves of Solanum tuberosum subsp. tuberosum cv. Bintje after treatment with culture filtrate (CF) from Erwinia carotovora subsp. carotovora strain SCC3193 (Pirhonen et al., 1988). Local treatment of potato leaves was done by applying 20-30 μl of CF to each leaf distributed in 4 to 6 different spots by gently pressing the tip of an automatic pipette against the leaf surface. (b) Accumulation of PRK transcripts in mini-tubers inoculated with 30-45 μl of CF applied by an automatic pipette. In (a) and (b), the amount of the corresponding RNA samples is indicated by a photo of the ethidium bromide-stained formaldehyde gels used for blotting. Potato plants used were grown axenically on MS medium (Murashige and Skoog 1962) for 3-4 weeks at 22° C. with a 14 hours light regime (100 to 150 μmol s⁻¹ m⁻²). In vitro plants grown for 3-4 weeks were either treated as indicated or transferred to soil and grown under gradually decreasing humidity but otherwise under similar conditions as indicated above for another ten days before treatment. Mini-tubers (1-3 grams fresh weight) were obtained from the soil plants grown under the same conditions for another 30-45 days. Three or more plants or mini-tubers were harvested after treatment at the indicated time points and total RNA was isolated (Verwoerd et al., 1989) and analyzed by RNA-gel blot experiments (Vidal et al., 1998). Each experiment was repeated twice or more. 10 μg of total RNA was used for each time point and hybridized with a 1 Kb PRK-4 probe corresponding to the extracellular domain, and labeled with [α-³²P]dCTP by random priming (Amersham International, UK).

[0031]FIG. 5 depicts the analysis of PRK mRNA accumulation in potato leaves after treatment with CF from E. carotovora subsp. carotovora and short oligogalacturonides. (a) Quantification of the PRK hybridization signals shown in b. The values shown are relative to the highest expression taken as 100%. Calculations were done using data obtained from Phosphor Imager (Fujifilm Bas-1500). (b) Accumulation of PRK mRNAs in potato leaves treated with 20-30 μl of the following: CF, 1 mM di-galacturonic acid (dimers) in water, 1 mM tri-galacturonic acid (trimers) in water, and H₂O which was used as a wound control. Dimers and trimers were purchased from Sigma (St. Louis, Mo.). The experimental conditions were otherwise as described in the legend to FIG. 4. (c) RT-PCR analysis of PRK-1 and 4. Leaf samples were treated as described in (b) and tuber samples as described in FIG. 4b. RT-PCR was performed as described by Sambrook and Russell (2001). For all samples, 1μg of total RNA (DNA-free) was reverse transcribed in a final volume of 50 μl. The resulting cDNA was amplified by PCR using 1 μl of the RT reaction in a final volume of 50 μl and the following cycling conditions: 94° C. for 4 min; (94° C. for 30 s, 62° C. for 60 s, 72° C. for 60 s) 3 cycles; (94° C. for 30 s, 60° C. for 60 s, 72° C. for 60 s) 35 cycles; elongation step at 72° C. for 5 min. The primers used were 5′-CCAACCATGGCAGCTGTTGTTCTC-3′ for PRK-1 and PRK-4; 5′-CACGTACACTAAAAGTGGTACCAACAC-3′ for PRK-1 and 5′-AAGAGGGGTACGGAAGGAGTTC-3′ for PRK-4. The RT reaction with all the components but reverse transcriptase or without RNA were used as controls and did not give any bands after PCR amplification (data not shown).

[0032]FIG. 6 depicts the deduced amino acid sequence of PRK-1 with 676 amino acids (SEQ ID:1).

[0033]FIG. 7 depicts the deduced amino acid sequence of PRK-2 with 676 amino acids (SEQ ID:2).

[0034]FIG. 8 depicts the deduced amino acid sequence of PRK-3 with 651 amino acids (SEQ ID:3).

[0035]FIG. 9 depicts the deduced amino acid sequence of PRK-4 with 676 amino acids (SEQ ID:4).

[0036]FIG. 10 depicts the nucleic acid sequence of PRK-1 cDNA with 2201 nucleotides, accession number AJ306626 (SEQ ID:5). The poly(A)-tail is not included in the 2201 nucleotides.

[0037]FIG. 11 depicts the nucleic acid sequence of PRK-2 cDNA with 2225 nucleotides, accession number AJ306627 (SEQ ID:6). The poly(A)-tail is not included in the 2225 nucleotides.

[0038]FIG. 12 depicts the nucleic acid sequence of PRK-3 cDNA with 2115 nucleotides, accession number AJ306628 (SEQ ID:7). The poly(A)-tail is not included in the 2115 nucleotides.

[0039]FIG. 13 the nucleic acid sequence of PRK-4 cDNA with 2387 nucleotides, accession number AJ306629 (SEQ ID:8). The poly(A)-tail is not included in the 2387 nucleotides.

[0040]FIG. 14 depicts the use Arabidopsis thaliana PRK. One full-length cDNA from the corresponding Arabidopsis gene has been isolated. The expression pattern of the Arabidopsis gene is similar to that of PRK responding to Erwinia carotovora.

[0041]FIG. 15 depicts the analysis of AtPRKs transgenic Arabidopsis plants. Transgenic Arabidopsis plants overexpressing this gene (sense) as well as transgenic plants where the expression of this gene is silenced (antisense) have been produced.

[0042]FIG. 16 depicts the sequence of Atpr3mia (SEQ ID:9), sequenced by the present inventors and differing from the corresponding sequence in the databank with accession number CAA 18465. It differs especially at the region of the transmembrane domain of the protein probably due to the computer programs used.

[0043]FIG. 17 depicts SEQ ID NO:10 (top) and SEQ ID NO:11 (bottom).

[0044]FIG. 18 depicts SEQ ID NO:12 (top) and SEQ ID NO:13 (bottom).

[0045]FIG. 19 depicts table of similarities of the extracellular (C-terminal) domain.

[0046]FIG. 20 depicts amino acid sequence of Arabidopsis Atmia prk (SEQ ID NO:14).

[0047]FIG. 21 depicts comparision between Atmia prk and databank sequence.

[0048]FIG. 22 depicts Atprks (Arabidopsis) ext and Stprks (Solanum tuberosum) extracellular cys similarity.

[0049]FIG. 23 depicts Atprks (Arabidopsis) ext and Stprks (Solanum tuberosum) extracellular cys similarity.

[0050]FIG. 24 depicts Atprks (Arabidopsis) ext and Stprks (Solanum tuberosum) extracellular cys similarity.

DETAILED DESCRIPTION OF THE INVENTION

[0051] Definitions

[0052] In the present invention the terms used have the meaning they generally have in the fields of molecular biology, recombinant DNA technology, botany and plant pathology. Some terms, however, are used with a somewhat deviating or broader meaning. Accordingly, in order to avoid uncertainty caused by terms with unclear meaning some of the terms used in the specification and in the claims are defined in more detail below.

[0053] The term “ligand” means any molecule, that binds tightly and specifically to a macromolecule, usually but not necessary a protein, forming a macromolecule-ligand complex, or to a receptor forming a receptor-ligand complex.

[0054] The term “elicitor” means molecules produced by the presence or action of pathogen or pest or mechanical damage that induce a response by the host.

[0055] The term “receptor” means any protein that binds a specific extracellular signaling molecule (ligand) and then initiates a cellular response. Receptors can be located within the cell or in the plasma membrane with their ligand-binding domain exposed to the external medium.

[0056] The term “pathogen” means, but is not limited to bacteria, viruses, nematodes, fungi or insects (see e.g. Agrios, Plant Pathology, Academic Press, San Diego, Calif., 1997).

[0057] The term “PRK” means potato receptor-like kinase.

[0058] The term “Atprk” means potato receptor-like kinase from Arabidopsis.

[0059] The term “potato receptor like kinase (PRK)-like compounds” means compounds, which act as PRK-like proteins. They include polypeptides “substantially homologous” at amino acid level having a significant similarity or identity of at least 60%, more preferred embodiments include at least, 65%, 70%, 75%, 80%, most preferably more than 85% with the reference sequence.

[0060] The term “PRK-like compounds” means protein molecules or polypeptides being substantially homologous to PRK at amino acid level. Said “PRK-like molecules” are obtainable by isolation from natural sources. The PRK-like molecules are also producible by synthetic, semisynthetic, enzymatic and other biochemical or chemical methods including recombinant DNA techniques.

[0061] The term “PRK-like compounds” also comprises polypeptides having the structure, properties and functions characteristic of PRK-like proteins, including PRK-like proteins, wherein one or more amino acid residues are substituted by another amino acid residue. Also truncated, complexed or chemically substituted, forms of said PRK-like proteins are included in the term. Chemically substituted forms include for example, alkylated, esterified, etherified or amidized forms with a low substitution degree, especially using small molecules, such as methyl or ethyl, as substituents, as long as the substitution does not disturb the properties and functions of the PRK-like proteins. The truncated, complexed and/or substituted variants of said polypeptides are producible by synthetic or semisynthetic, including enzymatic and recombinant DNA techniques. The only other prerequisite is that the derivatives still are substantially homologous with and have the properties and/or express the functions characteristic of PRK-like proteins.

[0062] The term “PRK-like compounds” otherwise covers all possible splice variants of potato receptor like kinase (PRK). The PRK-like compounds can exist in different isoforms or allelic forms.

[0063] More specifically “PRK-like proteins” are substantially homologous with the amino acid sequence SEQ ID:1, SEQ ID:2, SEQ ID:3, SEQ ID:4 or SEQ ID NO:9.

[0064] The term “isoform” refers to the one of several forms of the same protein, whose amino acid sequences differ slightly but whose general activity is identical. “Isoforms” may originate from different sources, e.g. different plant species. Isoforms of PRK compounds can be generated by the cleavage. Different enzymatic and non-enzymatic reactions, including proteolytic and non-proteolytic reactions, are capable of creating truncated, derivatized, complexed forms of PRK proteins.

[0065] In the present invention the term “PRK-like compounds” includes nucleic acid sequences, which belong to the active PRK-like compounds of the present invention and which comprise isolated or purified “nucleic acid sequences” encoding PRK-like proteins or nucleic acid sequences with substantial similarity. They can be used as such or introduced into suitable transformation or expression vectors, which in turn can be introduced into suitable host organism to provide procaryotic, eukaryotic organisms as well as transgenic plants capable of expressing altered levels of PRK-like proteins.

[0066] The term “nucleic acid sequences” refers to single of double-stranded polymer of deoxyribonucleotide or ribonucleotide bases. It includes chromosomal DNA, self-replicating plasmids, polymers of DNA or RNA. The “nucleic acid sequences” of the present invention are not in their natural state but are isolated and purified from their natural environment as transiently expressed mRNAs from a tissue. Thereafter the mRNAs are purified and multiplied in vitro in order to provide by technical means new copies, which are capable of encoding said PRK-like proteins. The nucleic acid sequences include both genomic sequences and cDNA.

[0067] The term “genomic sequence” means the corresponding sequence present in the nucleus of the plant cells and comprising introns as well as exons. In the present context the term “cDNA” means a DNA sequence obtainable by reversed translation of mRNA translated from the genomic DNA sequence including the complementary sequence.

[0068] The term “nucleic acid sequence encoding PRK or PRK-like proteins” means nucleic acid sequences encoding PRK or substantially homologous sequences. Said sequences or their complementary sequences or nucleic acid sequences containing said sequences or parts thereof, e.g. fragments truncated at the 3′-terminal or 5′-terminal end, as well as such sequences containing point mutations, are especially useful for as probes, primers and for preparing DNA constructs, plasmids and/or vectors useful for modulating the level of expression in plant tissues.

[0069] It is however clear for those skilled in the art that other nucleic acid sequences are capable of encoding PRK-like proteins and useful for their production can be prepared. Said nucleic acid sequences and/or their complementary sequences should be capable of hybridizing under highly stringent condition (Sambrook and Russell, 2001) with SEQ ID:5, SEQ ID:6, SEQ ID:7 or SEQ ID:8.

[0070] The nucleic acid sequences of the present invention should have a substantial similarity with the sequences encoding PRK or PRK-like proteins. “Substantial similarity” in this context means that the nucleotide sequences fulfill the prerequisites defined above and have a significant similarity, i.e. a sequence identity of at least of at least 40%, more preferred embodiments include at least 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, most preferably more than 85% with the reference sequence.

[0071] The term “nucleic acid sequences encoding PRK or PRK-like proteins” include their truncated or complexed forms as well as point mutations of said nucleic acid sequences as long as they are capable of encoding amino acid sequences having the essential structural features as well as the properties and/or functions of said PRK-like compounds.

[0072] The nucleic acid sequences are useful as such or inserted in transformation or expression vectors or host, said nucleic acid sequences being capable of encoding PRK or PRK-like proteins which are recognizable by binding substances specifically recognizing said PRK or PRK-like proteins. The nucleic acid sequences are useful in gene therapy or for preventing the genes causing the disease from expressing the gene products causing the diseases.

[0073] The PRK-like compounds include in addition to the proteins and nucleic acid sequences also binding substances.

[0074] The term “binding substances” means substances, which are capable of recognizing and specifically binding to natural PRK and/or PRK-like proteins or at least one specific portion of said molecules. Such binding substances are for example antibodies, receptors or ligands or proteins, specifically recognizing or binding to PRK or PRK-like proteins, ligands of PRK-like proteins or other binding proteins or peptides, comprising e.g. specific portions of said PRK-like compounds, but above all they mean antibodies capable of specifically recognizing one or more PRK-like compounds alone or in any combination. The antibodies include both polyclonal and/or monoclonal antibodies as well as fragments or derivatives thereof. Preferably, such binding substances, which recognize and bind to specific epitopes or active sites of the PRK-like compounds.

[0075] Said “binding substances” can be produced using specific domains of PRK-like compounds, their isomers as well as their fragments, derivatives and complexes with the prerequisite that they are capable of functioning in respective signaling pathway.

[0076] General Description of the Invention

[0077] Identification of potato genes responsive to cell wall-degrading enzymes of Erwinia carotovora resulted in isolation of cDNA clones for four related receptor-like protein kinases. One of the putative serine-threonine protein kinases might have arisen through alternative splicing. These potato receptor-like kinases (PRK1-4) were highly similar (91-99%) most likely constituting a family of related receptors. All PRKs and four other plant receptor like kinases (RLKs) share in their extracellular domain a conserved bi-modular pattern of cysteine repeats distinct from that in previously characterized plant RLKs, suggesting they represent a new class of receptors. The corresponding genes were rapidly induced by E. carotovora culture filtrate (CF) both in the leaves and tubers of potato. Furthermore, the genes were transiently induced by short oligogalacturonides. The structural identity of PRKs and their induction pattern suggested that they constitute part of the early response of potato to E. carotovora infection.

[0078] The present inventors were interested in understanding potato (Solanum tuberosum) defense responses against E. carotovora. In order to isolate potato genes, which are involved in defense during the early stages of the plant-pathogen interaction, plants were inoculated with E. carotovora subsp. carotovora strain SCC3193 (Pirhonen et al., 1988) and pathogen induced cDNA clones were isolated by suppression subtractive hybridization (SSH) (see Birch et al., 1999 for methods). One of the 25 characterized cDNAs corresponding to CF-induced genes predicted a polypeptide showing similarity to protein kinases and was analyzed further. First the full-length cDNA corresponding to the original 206-bp cDNA fragment was isolated. To achieve this a cDNA library was constructed with RNA samples from CF-treated leaves using the SMART™ RACE cDNA Amplification kit (Clonetech Laboratories, Inc.). Screening of this library resulted in isolation of four cDNAs with different EcoRI-restriction patterns (data not shown) all homologous to the 206-bp cDNA fragment.

[0079] The four full-length cDNAs were designated PRK-1 (2201 nucleotides, accession number AJ306626), PRK-2 (2225 nucleotides, accession number AJ306627), PRK-3 (2115 nucleotides, accession number AJ306628), and PRK-4 (2387 nucleotides, accession number AJ306629) for potato receptor-like protein kinase (PRK). Their predicted open reading frames encoded 676 amino acid polypeptides with a calculated molecular mass of 75 kDa for PRK-1, 2 and 4 and a 651 amino acid polypeptide with a calculated molecular mass of 72 kDa for PRK-3 (FIG. 1). A hydropathy plot analysis (Kyte and Doolittle, 1982) indicated that the PRKs have two very hydrophobic regions (FIG. 1); one at the amino terminus indicative of a signal peptide (von Heijne, 1990), followed by a 255-280 amino acids hydrophilic domain that contains 6-7 putative glycosylation sites, and a second hydrophobic region of 23 amino acids which is followed by basic residues indicative of Type I integral membrane proteins (Singer, 1990). Alignment of their deduced amino acid sequences and comparison with similar sequences from databases showed that they were related of their C-terminal domains to plant receptor-like protein kinases (RLK), (FIG. 2A). The C-terminal domains of the four PRKs contain all the 11 subdomains conserved among different kinases (Hanks et al., 1988) including the 15 invariant amino acids with the right organization (FIG. 2A). In addition, the motifs in the catalytic core, DLKXXN in subdomain VI and APE in sub domain VIII are indicative of serine-threonine protein kinases (Hanks and Quinn, 1991). Characterization of the potato genome by EcoRV digestion, which does not cut the PRK cDNAs followed by Southern hybridization to a PRK specific probe (FIG. 3) suggested that there are probably at least 3 genes in this family. Although it remains to be biochemically confirmed, these data strongly suggest that the four potato PRKs form a family of receptor-like serine-threonine protein kinases.

[0080] PRK-2, 3 and 4 exhibit 98-99% amino acid similarity while PRK-1 shows 91-93% similarity to PRK-2, 3 and 4 (FIG. 1). The difference between PRK- 1 and the other PRKs is accentuated when comparing the extracellular domains. The similarity of PRK-1 to PRK-2, 3 and 4 diminishes to 86-87% while the similarity between PRK-2, 3 and 4 is still 98-99%. Interestingly, PRK-3 presents a gap of 25 amino acids in a region of the putative extracellular domain that contains a conserved cysteine and a putative glycosylation site in the other PRKs (FIG. 1). Analysis of the corresponding cDNA sequences at this region revealed that the 25 amino acids difference of PRK-3 could be generated by alternative splicing from a different isoform (FIG. 2B). Identification of highly conserved sequences for splice sites (Brown and Simpson, 1998) flanking the 25 amino acids region strongly suggests the possibility for alternative splicing (FIG. 2B). This would result in removal of the codons for the 25 amino acids and as an additional consequence of such splicing to an amino acid substitution (serine instead of alanine) in PRK-3. Results of Southern analysis of the potato genome hybridized with a PRK probe specific to the fragment covering the putative intron supported this notion (FIG. 3). We could only detect a 700 bp HhaI and PstI fragment hybridizing to the probe but not a 625 bp fragment, which would be the expected size if the genomic DNA corresponding to PRK-3 would contain a deletion instead of an intron. Recently, it has been shown in Ipomea nil that alternative splicing occurred in a leucine-rich repeat receptor-like kinase (Bassett et al., 2000). Interestingly, a similar kind of splicing event was suggested in the extracellular domain of TGF-β type II receptors from mouse and human, which showed a 25 amino acids insertion containing one or two cysteines residues plus one putative glycosylation site, and one amino acid substitution at the splice junction (Hirai and Fujita, 1996; Suzuki et al., 1994). This splicing event resembles the one that can be predicted for the potato PRKs, suggesting that a similar mechanism could be involved in the processing of receptor-like serine-threonine protein kinases in different types of eukaryotic cells. This probably reflects the ability of cells to create receptors with the same function but different affinities for a ligand or structurally similar ligands.

[0081] Based on the structure of the putative extracellular domains, plant RLKs have been classified into several major classes (Braun and Walker, 1996; Walker, 1994). These include (i) the S-domain RLKs which contain extracellular domains homologous to the S-locus glycoproteins of Brassicaceae (Nasrallah et al., 1993; Walker and Zhang, 1990, Stein et al., 1991), (ii) the leucine rich repeat (LRR) RLKs such as Xa21 from rice, TMK1 and RLK5 from Arabidopsis (Chang et al., 1992; Song et al., 1995; Walker, 1993) and (iii) RLKs with the epidermal growth factor-like repeat (EGF) such as pro25 and the WAKs from Arabidopsis (He et al., 1999; Kohorn et al., 1992). Moreover, several RLKs with different types of extracellular domains have been identified recently (reviewed by Satterlee and Sussman, 1998). Comparison of PRKs with already known plant or other eukaryotic receptors, showed relation to PvPR20-1, a RLK of a new type from Phaseoulus vulgaris (Lange et al., 1999) and three different putative Arabidopsis RLKs (FIG. 2C). Alignment of their extracellular domains showed that: (i) the domains were 45 to 59% similar, i.e. 41 to 55% different, (ii) all of them contained 6-9 glycosylation sites except for one gene from Arabidopsis that had 4 glycosylation sites, (iii) the relative positions of 4 of the glycosylation sites were conserved and, (iv) all contained a conserved pattern of cysteine residues. This cysteine pattern presents two modules containing 6 cysteines each. The first module starts close to the putative signal peptide at the amino terminus and contains a C-X₍₄₉₋₅₃₎-C-X₍₈₎-C-X₍₂₎-C-X₍₁₁₎-C-X₍₁₂₋₁₄₎-C motif. It is followed by 75-77 amino acids that link it to the second module that contains a C-X₍₈₎-C-X₍₂₎-C-X₍₁₀₎-C-X₍₀₋₁₎-C-X₍₁₂₎-C motif, followed by a 42-46 amino acids segment before the putative transmembrane domain. Interestingly, PRK-3 lacks a cysteine in the first module while PRK-4 lacks a cysteine in the second module (FIG. 2C). Several eukaryotic receptors exhibit conserved cysteines, which could be involved in disulfide bond formation that may determine the general fold of the proteins. Furthermore, a similar cystine knot structure has been described in different families of animal receptor kinases (McDonald and Hendrickson, 1993; Sun and Davies, 1995). On the other hand, different plant RLKs contain cysteine patterns (Chen, 2001; He et al., 1999; Kohorn et al., 1992; Satterlee and Sussman, 1998; Walker, 1994), but we failed to find the cysteine pattern described here in the extracellular domain of those sequences. Recently, Chen (2001) described a superfamily including a number of Arabidopsis RLKs and other proteins with C-rich repeats. Interestingly, part of the bi-modular cysteine pattern in PRKs described above (-C-X₍₈₎-C-X₍₂₎-C-) is also found in this superfamily of proteins. In conclusion, the structural similarities described and especially the conserved bi-modular cysteine pattern shared by the PRKs, PvPR20-1 from Phaseoulus vulgaris (Lange et al., 1999) and three different putative Arabidopsis RLKs, suggest that they represent a new class of plant RLKs.

[0082] The expression of plant RLK genes have shown diverse patterns, while some of them displayed expression only in vegetative tissues (Kohorn et al., 1992), others were expressed only in reproductive tissues (Goring et al., 1992; Stein et al., 1991) and some have been shown in both vegetative and reproductive tissues (Pastuglia et al., 1997). Interestingly, some of the RLK genes are responsive to pathogens and elicitors, including PvRK20-1 from Phaseolus vulgaris (Lange et al., 1999), SFR2 from Brassica olearacea (Pastuglia et al., 1997), Wak1 (He et al., 1998) and RLKs (Du and Chen, 2000) from Arabidopsis thaliana and the disease resistance gene Xa21 from rice (Song et al., 1995). To elucidate the role of PRKs in plant response to E. carotovora we characterized the expression pattern of PRKs in different plant tissues after CF treatment of potato plants by RNA-gel blot hybridization (FIGS. 4a and b). The results show that leaf tissue treated locally with CF exhibits a fast accumulation of PRK transcripts with the highest level observed within one hour of treatment after which the level of mRNA decreased but stayed at elevated level up to 24 hours (FIG. 4a). The systemic leaves showed a very low and delayed induction of PRKs (FIG. 4a). A similar induction pattern to that of locally treated leaves was also observed in CF-treated potato mini-tubers (FIG. 4b). The early expression of PRK genes in response to CF-treatment strongly suggests that PRKs are involved in signal perception during potato defense responses against E. carotovora. Furthermore, the related structure and the related expression patterns of potato PRKs and PvRK20-1 suggest that these receptors could be involved in related cellular processes during the plant-pathogen interactions.

[0083] In plants very little is known about the nature of the ligands interacting with serine-threonine RLKs. Recently, it has been shown that the extracellular domain of a RLK from Arabidopsis, BRI1, perceives brassinosteroids (He et al., 2000). On the other hand, in animals it has been shown that the interleukin 2 and the epidermal growth factor receptors are up regulated by their own ligands (Clark et al., 1985; Deeper et al., 1985). In order to elucidate the nature of the inducer of potato PRKs, we characterized accumulation of the corresponding transcripts following treatment with di-oligogalacturonic acid and tri-oligogalacturonic acid (FIG. 5a and b), which have previously been shown to induce plant defense related genes responsive to E. carotovora (Norman et al., 1999). Plants treated with oligogalacturonides, showed a rapid but transient increase in PRK transcript levels while a very low induction was observed in water-treated wound control plants. This low but reproducible wound response (5 a and b) could have been caused by a release from the plant of short oligogalacturonide elicitors during the treatment. The PRK transcripts were induced to similar levels (5 to 10-fold) during the first hour by both oligogalacturonides and CF. However, there was a distinct difference in expression patterns between CF and oligogalacturonide-treated samples at later time points. In the CF-treated samples, the level of PRKs was reduced during the second hour and continue unchanged at four hours, while plants treated with oligogalacturonides showed a higher level of induction during the second hour that was drastically decreased to control levels at four hours. The difference on the kinetics of PRK transcripts accumulation between CF and oligogalacturonide-treated plants might reflect the fact that the former contains an enzymatic solution, which is releasing different types of cell-wall fragments during several hours as maceration proceeds (data not shown) while the latter is a solution with a fixed concentration of oligogalacturonides. On the other hand, the drastic decrease of PRK mRNA levels at four hours of oligogalacturonide treatment may indicate the involvement of a different type of signal that controls the temporal regulation of PRK expression levels. RT-PCR was used to elucidate whether the different PRKs exhibited differences in their expression patterns (FIG. 5c). Due to extensive sequence similarities we could unambiguously distinguish only between PRK-1 and 4 but not between PRK-2 and 4. The results indicate that both genes PRK-1 and 4 are expressed similarly in response to CF and short oligogalacturonides in both leaf and tuber tissues, although we can not rule out the possibility of small differences in their expression patterns. PRK-3 specific PCR products were not detected suggesting a low level of expression.

[0084] In conclusion, the results of the expression studies demonstrate that short oligogalacturonides act as elicitors of PRK expression and indicate that E. carotovora released oligogalacturonides play an important role eliciting PRKs during the early stage of the potato-Erwinia interaction. Furthermore, the results suggest that the PRKs are involved in perception of E. carotovora by the host plant.

[0085] One full-length cDNA Atpr3mia (SEQ ID:9) from the corresponding Arabidopsis gene was isolated. It differs from the corresponding sequence in the databank with accession number CAA18465. It differs especially at the region of the transmembrane domain of the protein probably due to the computer programs used. The expression pattern of the Arabidopsis gene is similar to that of PRK responding to Erwinia carotovora. Transgenic Arabidopsis plants overexpressing this gene (sense) as well as transgenic plants where the expression of this gene is silenced (antisense) (FIG. 15) have been produced.

[0086] The receptors of the present invention may also participate in other functions than conferring disease resistance for example regulation of growth in different plant species.

Example 1 Atmia prk Transgenic Arabidopsis Infected with Erwinia carotovora subsp. carotovora SCC1

[0087]Arabidopsis thaliana plants were transformed using the Agrobacterium-mediated transformation method of Clough and Bent (1999, Plant Journal 16, 735-743). The Arabidopsis plants were transformed using vacuum infiltration method without plant tissue culture or regeneration. Developing floral tissues were dipped into a solution containing Agrobacterium tumefaciens, 5% sucrose and 500 microliters per litre of surfactant Silwet L-77. Plant tissue culture media, the hormone benzylamino purine and pH adjustment were unnecessary.

[0088] Arabidopsis plants were transformed with Atmia prk gene (SEQ ID NO:9) and they were subsequently infected with Erwinia carotovora subsp. carotovora SCC1. 3-week old seedlings of Arabidopsis transgenic lines where the Arabidopsis homolog of PRK (Atmia prk) is overexpressed (S 2142, S 1713) or silenced with antisense constructs (AS 1961, AS 1564) were used. The number of plants exhibiting disease symptoms over those locally inoculated with different size of inocula of the Erwinia strain SCC 1 are presented in Table 1. TABLE 1 Plants exhibiting disease symptoms (at 96 hrs)/total of treated plants SCC1 inoculum. S 2142 S 1713 AS 1961 AS 1564 PDE control 50.000 CFU 1/16 1/12 9/24 7/17 3/17 20.000 CFU 1/18 nt 5/18 9/18 5/18  7.500 CFU 2/18 1/12 8/12 5/17 3/12

[0089] The PDE control indicates transgenic Arabidopsis harboring the transformation vector without any PRK insert. The plants were locally inoculated without wounding and the development of disease symptoms assessed 96 hrs post inoculation.

[0090] The results demonstrate that plants overexpressing the Atmia prk show enhanced disease resistance [only 4/52 (S 2142) and 2/24 (S 1713) plants with disease symptoms] when compared to the vector PDE control (11/47 plants with disease symptoms). In contrast, plants where the gene is silenced by antisense constructs are more sensitive to the pathogen (22/54 (AS 1961) and 21/52 (AS 1564) plants with disease symptoms.

[0091] In conclusion, the results indicate, that the Atmia prk gene is required for full disease resistance in Arabidopsis. The results also indicate that overexpression of the Atmia prk gene enhances disease resistance.

[0092] The overexpression of Atmia prk gene or genes can be used to enhance disease resistance in various agricultural plants, in particular disease resistance to necrotrophic pathogens and possibly herbivores. Another application could be additional enhancement of the resistance of such transgenic lines by adding chemical preparations (e.g. by spraying fields) containing oligouronides. The above mentioned preparations might also be used to enhance resistance of normal (not transformed with prk) plants.

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1 14 1 676 PRT Solanum tuberosum PRK-1 1 Met Ala Ile Gln Lys Trp Leu Leu Phe Leu Phe Leu His Leu His Val 1 5 10 15 Leu Asn Ile Val Ala Gln Leu Pro Asp Leu Arg Phe Gly Ile Cys Gly 20 25 30 Lys Ser Gly Asn Tyr Thr Glu Asn Ser Thr Tyr Lys Asn Asp Leu Asn 35 40 45 Thr Leu Leu Thr Ser Leu Ser Ser Lys Ile Asp Lys Tyr Gly Phe Tyr 50 55 60 Asn Ala Ser Ile Gly Gln Asn Ser Asp Arg Ala Ser Val Ile Val Leu 65 70 75 80 Cys Arg Gly Asp Val Glu Leu Asp Asp Cys Arg Gly Cys Val Asp Asn 85 90 95 Val Val Gln Lys Ile Ala Gln Leu Cys Pro Asn Gln Lys Glu Val Phe 100 105 110 Gly Gly Tyr Asp Gly Cys Met Leu Gln Tyr Ser Asn Gln Ser Ile Ile 115 120 125 Asp Thr Pro Ser Leu Ser Val Gln Leu Phe Leu Trp Asn Thr Ala Asn 130 135 140 Ala Ser Lys Pro Glu Glu Phe Asn Gln Glu Leu Gly Lys Leu Leu Glu 145 150 155 160 Asn Leu Arg Asp Arg Ala Ala Gln Gly Gly Pro Leu Gln Lys Tyr Ala 165 170 175 Thr Gly Thr Thr Ile Gly Pro Asp Ile Gln Pro Ile Tyr Ala Leu Val 180 185 190 Gln Cys Thr Pro Asp Leu Ser Arg Gln Ser Cys Phe Asp Cys Leu Thr 195 200 205 Asp Ala Tyr Gly Thr Leu Pro Gln Cys Pro Cys Leu Gly Lys Thr Gly 210 215 220 Gly Arg Ile Ile Gly Ile Arg Cys Asn Phe Arg Tyr Glu Ile Ser Arg 225 230 235 240 Phe Phe Val Asp Val Pro Leu Glu Ala Pro Pro Pro Ala Gly Asn Asp 245 250 255 Asn Lys Thr Val Pro Thr Gly Thr Glu Asn Lys Thr Pro Pro Thr Gly 260 265 270 Lys Asp Asp Lys Thr Thr Arg Thr Ile Ile Ile Ile Val Val Ser Thr 275 280 285 Val Thr Ile Val Ile Leu Met Ile Cys Ile Ala Val Ile Leu Ile Arg 290 295 300 Arg Arg Lys Arg Lys Leu Val Asn Gly Ile Gln Gly Thr Ser Val Asp 305 310 315 320 Asp Thr Ser Ile Ala Glu Ser Phe Gln Tyr Asp Phe Ser Ala Ile Arg 325 330 335 Ala Ala Thr Asp Asp Phe Ser Asp Ala Asn Lys Leu Gly Glu Gly Gly 340 345 350 Phe Gly Pro Val Tyr Lys Gly Lys Leu Gln Asn Gly Gln Glu Val Ala 355 360 365 Val Lys Arg Leu Ser Ala Asp Ser Gly Gln Gly Asp Leu Glu Ser Lys 370 375 380 Asn Glu Val Leu Leu Val Ala Arg Leu Gln His Arg Asn Leu Val Arg 385 390 395 400 Leu Leu Gly Phe Cys Leu Asp Gly Thr Glu Arg Leu Leu Val Tyr Glu 405 410 415 Phe Val Pro Asn Ala Ser Leu Asp His Phe Leu Phe Asp Ser Val Lys 420 425 430 Arg Arg Gln Leu Asp Trp Glu Arg Arg Ser Lys Ile Ile Gly Gly Ile 435 440 445 Ala Lys Gly Ile Leu Tyr Leu His Glu Asp Ser Arg Leu Arg Ile Ile 450 455 460 His Arg Asp Leu Lys Ala Ser Asn Val Leu Leu Asp Ala Glu Met Asn 465 470 475 480 Pro Lys Ile Ser Asp Phe Gly Met Ala Arg Leu Phe Glu Leu Asp Glu 485 490 495 Thr Gln Gly Ser Thr Asn Arg Ile Val Gly Thr Tyr Gly Tyr Met Ala 500 505 510 Pro Glu Tyr Ala Met His Gly Gln Phe Ser Val Lys Ser Asp Val Phe 515 520 525 Ser Phe Gly Val Leu Val Leu Glu Ile Leu Ser Gly Gln Lys Asn Thr 530 535 540 Cys Phe Arg Asn Gly Glu Ser Val Glu Asp Leu Leu Ser Phe Ala Trp 545 550 555 560 Leu Ser Trp Arg Asn Gly Thr Thr Ile Asp Phe Val Asp Pro Met Leu 565 570 575 Lys Glu Ser Thr Gly Leu Ile Arg Asp Ile Met Arg Asn Ile His Ile 580 585 590 Ala Leu Leu Cys Val Gln Glu Ser Val Ala Asp Arg Pro Thr Met Ala 595 600 605 Ala Val Val Leu Met Leu Ser Ser Phe Ser Leu Ser Leu Pro Met Pro 610 615 620 Ser Gly Pro Ala Phe Tyr Met His Ser Asn Ile Thr Ala Glu Thr Ser 625 630 635 640 Leu Ile Lys Glu Tyr Asn Thr Arg Met Thr Asp Ser Ser Glu Leu Ala 645 650 655 Lys Ser Lys Ser Ile Gly Ser Ser Arg Asn Glu Ala Ser Ile Ser Glu 660 665 670 Leu Tyr Pro Arg 675 2 676 PRT Solanum tuberosum PRK-2 2 Met Ala Ile Gln Lys Trp Leu Leu Ile Leu Phe Leu His Ile His Val 1 5 10 15 Leu Asn Ile Val Ala Gln Leu Pro Asp Leu Arg Phe Gly Ser Cys Gly 20 25 30 Gly Asn Gly Asn Tyr Thr Glu Asn Ser Thr Tyr Lys Asn Asn Leu Asn 35 40 45 Thr Leu Leu Thr Ser Leu Ser Ser Lys Ile Asp Asn Tyr Gly Phe Tyr 50 55 60 Asn Ala Ser Ile Gly Gln Asn Ser Asp Arg Ala Ser Val Ile Val Leu 65 70 75 80 Cys Arg Gly Asp Val Glu Leu Ala Asp Cys Arg Gly Cys Val Asp Asn 85 90 95 Val Val Gln Lys Ile Ala Gln Leu Cys Pro Asn Gln Lys Glu Val Phe 100 105 110 Gly Gly Tyr Asp Gly Cys Met Leu Gln Tyr Ser Asn Gln Ser Ile Leu 115 120 125 Glu Thr Thr Ser Phe Ser Leu Lys Tyr Tyr Phe Trp Asn Pro Ala Asn 130 135 140 Ala Thr Lys Pro Gly Glu Phe Asn Gln Glu Leu Gly Arg Leu Leu Glu 145 150 155 160 Asn Leu Arg Asp Arg Ala Val Asp Asp Gly Pro Leu Gln Lys Tyr Ala 165 170 175 Ser Gly Asn Ala Thr Gly Pro Asp Phe Gln Ala Ile Tyr Ala Leu Val 180 185 190 Gln Cys Thr Pro Asp Leu Ser Arg Gln Ser Cys Phe Ser Cys Leu Ser 195 200 205 Asp Ala Tyr Gly Asn Met Pro Arg Cys Pro Cys Leu Gly Lys Arg Gly 210 215 220 Gly Arg Ile Ile Gly Val Arg Cys Asn Phe Arg Tyr Glu Ser Ser Arg 225 230 235 240 Phe Phe Glu Asp Val Pro Leu Glu Ala Pro Pro Pro Ala Gly Asn Asp 245 250 255 Asn Thr Thr Val Pro Thr Gly Thr Asp Asp Lys Thr Val Pro Thr Gly 260 265 270 Glu Asp Asp Lys Thr Thr Arg Thr Ile Ile Ile Ile Val Val Ser Thr 275 280 285 Val Thr Val Val Ile Leu Ile Ile Cys Ile Ala Val Ile Leu Ile Arg 290 295 300 Arg Arg Lys Arg Lys Leu Val Asn Glu Ile Gln Ser Thr Ser Val Asp 305 310 315 320 Asp Thr Ser Ile Ala Glu Ser Phe Gln Tyr Asp Phe Ser Ala Ile Arg 325 330 335 Ala Ala Thr Asp Asp Phe Ser Asp Ala Asn Lys Leu Gly Glu Gly Gly 340 345 350 Phe Gly Pro Val Tyr Lys Gly Lys Leu Gln Asn Gly Gln Glu Val Ala 355 360 365 Val Lys Arg Leu Ser Ala Asp Ser Gly Gln Gly Asp Leu Glu Phe Lys 370 375 380 Asn Glu Val Leu Leu Val Ala Arg Leu Gln His Arg Asn Leu Val Arg 385 390 395 400 Leu Leu Gly Phe Cys Leu Asp Gly Thr Glu Arg Leu Leu Val Tyr Glu 405 410 415 Phe Val Pro Asn Ala Ser Leu Asp His Phe Leu Phe Asp Ser Val Lys 420 425 430 Arg Arg Gln Leu Asp Trp Glu Arg Arg Ser Lys Ile Ile Gly Gly Ile 435 440 445 Ala Lys Gly Ile Leu Tyr Leu His Glu Asp Ser Arg Leu Arg Ile Ile 450 455 460 His Arg Asp Leu Lys Ala Ser Asn Val Leu Leu Asp Ala Glu Met Asn 465 470 475 480 Pro Lys Ile Ser Asp Phe Gly Met Ala Arg Leu Phe Glu Leu Asp Glu 485 490 495 Thr Gln Gly Ser Thr Asn Arg Ile Val Gly Thr Tyr Gly Tyr Met Ala 500 505 510 Pro Glu Tyr Ala Met His Gly Gln Phe Ser Val Lys Ser Asp Val Phe 515 520 525 Ser Phe Gly Val Leu Val Leu Glu Ile Leu Ser Gly Gln Lys Asn Thr 530 535 540 Cys Phe Arg Asn Gly Glu Ser Val Glu Asp Leu Leu Ser Phe Ala Trp 545 550 555 560 Ser Ser Trp Arg Asn Gly Thr Thr Ile Asn Phe Val Asp Pro Met Leu 565 570 575 Lys Glu Ser Thr Gly Leu Ile Arg Asp Ile Met Arg Asn Ile His Ile 580 585 590 Ala Leu Leu Cys Val Gln Glu Ser Val Ala Asp Arg Pro Thr Met Ala 595 600 605 Ala Val Val Leu Met Leu Ser Ser Phe Ser Leu Ser Leu Pro Met Pro 610 615 620 Ser Gly Pro Ala Phe Tyr Met His Ser Asn Ile Thr Ala Gly Thr Ser 625 630 635 640 Leu Ile Gln Glu Tyr Asn Thr Arg Val Thr Asp Ser Ser Glu Arg Ala 645 650 655 Lys Ser Lys Ser Ile Gly Ser Ser Arg Asn Glu Ala Ser Ile Thr Glu 660 665 670 Leu Tyr Pro Arg 675 3 651 PRT Solanum tuberosum PRK-3 3 Met Ala Ile Gln Lys Trp Leu Leu Ile Leu Phe Leu His Ile His Val 1 5 10 15 Leu Asn Ile Val Ala Gln Leu Pro Asp Leu Arg Phe Gly Ser Cys Gly 20 25 30 Gly Asn Gly Asn Tyr Thr Glu Asn Ser Thr Tyr Lys Asn Asn Leu Asn 35 40 45 Thr Leu Leu Thr Ser Leu Ser Ser Lys Ile Asp Asn Tyr Gly Phe Tyr 50 55 60 Asn Ala Ser Ile Gly Gln Asn Ser Asp Arg Ala Ser Val Ile Val Leu 65 70 75 80 Cys Arg Gly Asp Val Glu Leu Ala Asp Cys Arg Gly Cys Val Asp Asn 85 90 95 Val Val Gln Lys Ile Ala Gln Leu Cys Pro Asn Gln Lys Glu Val Phe 100 105 110 Gly Gly Tyr Asp Gly Ser Asn Ala Thr Lys Pro Glu Glu Phe Asn Gln 115 120 125 Glu Leu Gly Arg Leu Leu Glu Asn Leu Arg Asp Arg Ala Val Asp Asp 130 135 140 Gly Pro Leu Gln Lys Tyr Ala Ser Gly Asn Ala Thr Gly Pro Asp Phe 145 150 155 160 Gln Ala Ile Tyr Ala Leu Val Gln Cys Thr Pro Asp Leu Ser Arg Gln 165 170 175 Ser Cys Phe Ser Cys Leu Ser Asp Ala Tyr Gly Asn Met Pro Arg Cys 180 185 190 Pro Cys Leu Gly Lys Arg Gly Gly Arg Ile Ile Gly Val Arg Cys Asn 195 200 205 Phe Arg Tyr Glu Ser Ser Arg Phe Phe Glu Asp Val Pro Leu Glu Ala 210 215 220 Pro Pro Pro Ala Gly Asn Asp Asn Thr Thr Val Pro Thr Gly Thr Asp 225 230 235 240 Asp Lys Thr Val Pro Thr Gly Glu Asp Asp Lys Thr Thr Arg Thr Ile 245 250 255 Ile Ile Ile Val Val Ser Thr Val Thr Val Val Ile Leu Ile Ile Cys 260 265 270 Ile Ala Val Ile Leu Ile Arg Arg Arg Lys Arg Lys Leu Val Asn Glu 275 280 285 Ile Gln Ser Thr Ser Val Asp Asp Thr Ser Ile Ala Glu Ser Phe Gln 290 295 300 Tyr Asp Phe Ser Ala Ile Arg Ala Ala Thr Asp Asp Phe Ser Asp Ala 305 310 315 320 Asn Lys Leu Gly Glu Gly Gly Phe Gly Pro Val Tyr Lys Gly Lys Leu 325 330 335 Gln Asn Gly Gln Glu Val Ala Val Lys Arg Leu Ser Ala Asp Ser Gly 340 345 350 Gln Gly Asp Leu Glu Phe Lys Asn Glu Val Leu Leu Val Ala Arg Leu 355 360 365 Gln His Arg Asn Leu Val Arg Leu Leu Gly Phe Cys Leu Asp Gly Thr 370 375 380 Glu Arg Leu Leu Val Tyr Glu Phe Val Pro Asn Ala Ser Leu Asp His 385 390 395 400 Phe Leu Phe Asp Ser Val Lys Arg Arg Gln Leu Asp Trp Glu Arg Arg 405 410 415 Ser Lys Ile Ile Gly Gly Ile Ala Lys Gly Ile Leu Tyr Leu His Glu 420 425 430 Asp Ser Arg Leu Arg Ile Ile His Arg Asp Leu Lys Ala Ser Asn Val 435 440 445 Leu Leu Asp Ala Glu Met Asn Pro Lys Ile Ser Asp Phe Gly Met Ala 450 455 460 Arg Leu Phe Glu Leu Asp Glu Thr Gln Gly Ser Thr Asn Arg Ile Val 465 470 475 480 Gly Thr Tyr Gly Tyr Met Ala Pro Glu Tyr Ala Met His Gly Gln Phe 485 490 495 Ser Val Lys Ser Asp Val Phe Ser Phe Gly Val Leu Val Leu Glu Ile 500 505 510 Leu Ser Gly Gln Lys Asn Thr Cys Phe Arg Asn Gly Glu Ser Val Glu 515 520 525 Asp Leu Leu Ser Phe Ala Trp Ser Ser Trp Arg Asn Gly Thr Thr Ile 530 535 540 Asn Phe Val Asp Pro Met Leu Lys Glu Ser Thr Gly Leu Ile Arg Asp 545 550 555 560 Ile Met Arg Asn Ile His Ile Ala Leu Leu Cys Val Gln Glu Ser Val 565 570 575 Ala Asp Arg Pro Thr Met Ala Ala Val Val Leu Met Leu Ser Ser Phe 580 585 590 Ser Leu Ser Leu Pro Met Pro Ser Gly Pro Ala Phe Tyr Met His Ser 595 600 605 Asn Ile Thr Ala Gly Thr Ser Leu Ile Gln Glu Tyr Asn Thr Arg Val 610 615 620 Thr Asp Ser Ser Glu Arg Ala Lys Ser Lys Ser Ile Gly Ser Ser Arg 625 630 635 640 Asn Glu Ala Ser Ile Thr Glu Leu Tyr Pro Arg 645 650 4 676 PRT Solanum tuberosum PRK-4 4 Met Ala Ile Gln Lys Trp Leu Leu Ile Leu Phe Leu His Ile His Val 1 5 10 15 Leu Asn Ile Val Ala Gln Leu Pro Asp Leu Arg Phe Gly Ser Cys Gly 20 25 30 Gly Asn Gly Asn Tyr Thr Glu Asn Ser Thr Tyr Lys Asn Asn Leu Asn 35 40 45 Thr Leu Leu Thr Ser Leu Ser Ser Lys Ile Asp Asn Tyr Gly Phe Tyr 50 55 60 Asn Ala Ser Ile Gly Gln Asn Ser Asp Arg Ala Ser Val Ile Val Leu 65 70 75 80 Cys Arg Gly Asp Val Glu Leu Ala Asp Cys Arg Gly Cys Val Asp Asn 85 90 95 Val Val Gln Lys Ile Ala Gln Leu Cys Pro Asn Gln Lys Glu Val Phe 100 105 110 Gly Gly Tyr Asp Gly Cys Met Leu Gln Tyr Ser Asn Gln Ser Ile Leu 115 120 125 Glu Thr Thr Ser Phe Ser Leu Lys Tyr Tyr Phe Trp Asn Pro Ala Asn 130 135 140 Ala Thr Lys Pro Glu Glu Phe Asn Gln Glu Leu Gly Arg Leu Leu Glu 145 150 155 160 Asn Leu Arg Asp Arg Ala Val Asp Asp Gly Pro Leu Gln Lys Tyr Ala 165 170 175 Ser Gly Asn Ala Thr Gly Pro Asp Phe Gln Ala Ile Tyr Ala Leu Val 180 185 190 Gln Cys Thr Pro Asp Leu Ser Arg Gln Ser Cys Phe Ser Cys Leu Ser 195 200 205 Asp Ala Tyr Gly Asn Met Pro Arg Arg Pro Cys Leu Gly Lys Arg Gly 210 215 220 Gly Arg Ile Ile Gly Val Arg Cys Asn Phe Arg Tyr Glu Ser Ser Arg 225 230 235 240 Phe Phe Glu Asp Val Pro Leu Glu Ala Pro Pro Pro Ala Gly Asn Asp 245 250 255 Asn Thr Thr Val Pro Thr Gly Thr Asp Asp Lys Thr Val Pro Thr Gly 260 265 270 Glu Asp Asp Lys Thr Thr Arg Thr Ile Ile Ile Ile Val Val Ser Thr 275 280 285 Val Thr Val Val Ile Leu Ile Ile Cys Ile Ala Val Ile Leu Ile Arg 290 295 300 Arg Arg Lys Arg Lys Leu Val Asn Glu Ile Gln Ser Thr Ser Val Asp 305 310 315 320 Asp Thr Ser Ile Ala Glu Ser Phe Gln Tyr Asp Phe Ser Ala Ile Arg 325 330 335 Ala Ala Thr Asp Asp Phe Ser Asp Ala Asn Lys Leu Gly Glu Gly Gly 340 345 350 Phe Gly Pro Val Tyr Lys Gly Lys Leu Gln Asn Gly Gln Glu Val Ala 355 360 365 Val Lys Arg Leu Ser Ala Asp Ser Gly Gln Gly Asp Leu Glu Phe Lys 370 375 380 Asn Glu Val Leu Leu Val Ala Arg Leu Gln His Arg Asn Leu Val Arg 385 390 395 400 Leu Leu Gly Phe Cys Leu Asp Gly Thr Glu Arg Leu Leu Val Tyr Glu 405 410 415 Phe Val Pro Asn Ala Ser Leu Asp His Phe Leu Phe Asp Ser Val Lys 420 425 430 Arg Arg Gln Leu Asp Trp Glu Arg Arg Ser Lys Ile Ile Gly Gly Ile 435 440 445 Ala Lys Gly Ile Leu Tyr Leu His Glu Asp Ser Arg Leu Arg Ile Ile 450 455 460 His Arg Asp Leu Lys Ala Ser Asn Val Leu Leu Asp Ala Glu Met Asn 465 470 475 480 Pro Lys Ile Ser Asp Phe Gly Met Ala Arg Leu Phe Glu Leu Asp Glu 485 490 495 Thr Gln Gly Ser Thr Asn Arg Ile Val Gly Thr Tyr Gly Tyr Met Ala 500 505 510 Pro Glu Tyr Ala Met His Gly Gln Phe Ser Val Lys Ser Asp Val Phe 515 520 525 Ser Phe Gly Val Leu Val Leu Glu Ile Leu Ser Gly Gln Lys Asn Thr 530 535 540 Cys Phe Arg Asn Gly Glu Ser Val Glu Asp Leu Leu Ser Phe Ala Trp 545 550 555 560 Ser Ser Trp Arg Asn Gly Thr Thr Ile Asn Phe Val Asp Pro Met Leu 565 570 575 Lys Glu Ser Thr Gly Leu Ile Arg Asp Ile Met Arg Asn Ile His Ile 580 585 590 Ala Leu Leu Cys Val Gln Glu Ser Val Ala Asp Arg Pro Thr Met Ala 595 600 605 Ala Val Val Leu Met Leu Ser Ser Phe Ser Leu Ser Leu Pro Met Pro 610 615 620 Ser Gly Pro Ala Phe Tyr Met His Ser Asn Ile Thr Ala Gly Thr Ser 625 630 635 640 Leu Ile Gln Glu Tyr Asn Thr Arg Val Thr Asp Ser Ser Glu Arg Val 645 650 655 Lys Ser Lys Ser Ile Gly Ser Ser Arg Asn Glu Ala Ser Ile Thr Glu 660 665 670 Leu Tyr Pro Arg 675 5 2227 DNA Solanum tuberosum PRK-1 cDNA 5 atcaaaactt gcttgtaact taaacagaca tagccatggc tattcaaaag tggttactct 60 ttctgttttt gcatttacat gttctcaata ttgtagcgca gctgcctgat ttacgattcg 120 gtatatgtgg taagagtggt aactatactg agaatagtac ctacaagaac gatctaaaca 180 cactcctcac ttccctttcc tcaaaaatag ataagtatgg tttctacaat gcttctattg 240 gccaaaactc tgatagagcc agcgttattg tgttatgtag aggagatgtg gagctagacg 300 attgtcgcgg ttgtgtcgat aatgttgttc aaaagattgc acagttatgt cctaaccaaa 360 aagaagtttt cggtggctat gatggatgta tgttacagta ttcgaatcaa tccattatag 420 atactccatc gttgtctgtt caattatttc tttggaatac tgcgaatgcc tcaaaacccg 480 aggaatttaa ccaggagcta ggaaaattat tggaaaattt acgagaccgt gctgcacagg 540 gtggtcctct tcaaaaatat gctactggta ctacgatagg tccagatatt cagcctatat 600 atgcacttgt gcagtgtact cctgatttat ctcgtcagag ttgcttcgat tgcttaactg 660 acgcttatgg aaccttgcct caatgtccct gcctgggaaa gaccggtgga agaatcatag 720 ggattaggtg caacttccgt tatgaaattt cccgtttctt cgtcgatgtg ccgttggaag 780 ctccgccacc tgcaggaaat gataataaaa cagttccaac agggacggaa aataaaacac 840 ctccaactgg gaaggacgat aaaacaacac gaacaattat cattatcgtt gtgtcaactg 900 ttacaattgt tattcttatg atttgtattg ctgtcatctt gataaggagg cgaaagagga 960 aactggtgaa cggaattcag ggtacatctg tagatgatac tagtattgca gaatcttttc 1020 aatatgattt ttcggcaatt agagcagcaa cagatgactt ctcagatgct aataagcttg 1080 gagaaggtgg atttggtcct gtgtacaagg gtaagcttca aaatggacaa gaagtagcag 1140 tgaaaaggtt atcagcagat tcaggccaag gtgatctaga atccaaaaat gaggtcttgt 1200 tggttgccag gcttcaacac aggaatttgg ttaggttgct gggattttgc ctagacggaa 1260 cagagcgact tcttgtctat gagtttgttc ccaatgcaag tcttgaccac ttcttatttg 1320 attcagttaa acgtaggcaa ttggattggg aaaggcgatc caaaatcata ggaggcattg 1380 ctaagggaat tctttatctt catgaggatt ctaggcttcg gatcattcac cgtgatctca 1440 aagctagtaa tgttctacta gatgcagaaa tgaatcctaa aatctcagat tttggcatgg 1500 caaggctatt tgaattagat gaaactcaag gcagcacaaa cagaattgtt gggacctatg 1560 gatatatggc accagagtat gcaatgcacg ggcaattttc cgtaaagtca gatgttttta 1620 gctttggagt actagtctta gaaattttaa gtggccaaaa aaacacttgt ttcagaaatg 1680 gagaatcggt ggaagacctt ttgagttttg cttggttaag ctggcgtaat ggaacaacta 1740 tagattttgt agatccaatg ctgaaggaaa gcacaggact gattcgtgac ataatgagaa 1800 acattcacat agctttattg tgcgttcaag aaagtgtggc tgatagacca accatggcag 1860 ctgttgttct catgctcagt agcttttcgt tgagtcttcc aatgccttca gggccagcat 1920 tctatatgca cagtaatatt accgcagaga cgtcgcttat taaagaatac aacacaagaa 1980 tgacagactc tagtgagcta gccaaaagta aatctattgg ttcatcacga aatgaggcat 2040 ccatatctga gttatatcct cgttaacatc cctagtatga taaccttttt ctctgagttt 2100 acttatttta tgtgttggta ccacttttag tgtacgtgta gtctagctaa caataattat 2160 tataaagttt atatgtaata gtaacactct ttttaataca gaaaaaaaaa aaaaaaaaaa 2220 aaaaaaa 2227 6 2242 DNA Solanum tuberosum PRK-2 cDNA 6 atcaaaactt gcttgtaact taaacagaca tagccatggc tattcaaaag tggctactca 60 ttctgttttt gcatatacat gttctcaata ttgtagcgca gctgcctgat ttacgatttg 120 gttcatgtgg tggcaatggt aactatactg agaatagtac ctacaagaac aatctaaaca 180 cactcctcac ttccctttcc tcaaaaatag acaactatgg tttctacaat gcttctattg 240 gtcaaaactc tgatagagcc agcgttattg tgttatgtag aggagatgtg gagctagccg 300 attgtcgcgg ttgtgtcgat aatgttgttc aaaagattgc acagttatgt cctaaccaaa 360 aagaagtttt cggtggctat gatggatgta tgttacagta ttcaaatcaa tccattttag 420 aaactacatc attttctctt aaatattact tttggaatcc agcgaatgcc acaaaacccg 480 gggaatttaa ccaggagcta gggagattat tggaaaactt acgagatcgt gctgtagatg 540 atggtcctct tcaaaaatat gctagtggta atgcgacagg tccagatttt caggctatat 600 atgcacttgt gcagtgcact cctgatttat ctcgtcagag ttgcttcagt tgcttaagtg 660 acgcttatgg aaacatgcct cgctgtccct gcctgggtaa gaggggtgga agaatcatag 720 gggttaggtg caacttccgt tatgaaagtt cccgtttctt cgaggatgtg ccgttggaag 780 ctccaccacc tgcaggaaat gataatacaa cagttccaac agggacggac gataaaacag 840 ttccaacagg agaggatgac aaaacaacac gaacaatcat cattatcgtt gtgtcaactg 900 ttacagttgt tattcttatt atttgtattg ctgtcatctt gataaggagg cgaaagagga 960 agctggtgaa cgaaattcag agtacatctg tagatgatac tagtattgca gaatcttttc 1020 aatatgattt ttcggcaatt agagcagcaa cagatgactt ctcagatgct aataagctcg 1080 gagaaggcgg atttggtcct gtgtacaagg gtaagcttca aaatggacaa gaagtagcag 1140 tgaaaaggtt atcagcagat tcaggccaag gtgatctaga attcaaaaat gaggtcttgt 1200 tggttgccag gcttcaacac aggaatttgg ttaggttgct gggattttgc ctagacggaa 1260 cagagcgact tcttgtctat gagtttgttc ccaatgcaag tcttgaccac ttcttatttg 1320 attcagttaa acgtaggcaa ttggattggg aaaggcgatc caaaatcata ggaggcattg 1380 ctaagggaat tctttatctt catgaggatt ctaggcttcg gatcattcac cgtgatctca 1440 aagctagtaa tgttctacta gatgcagaaa tgaatcctaa aatctcagat tttggcatgg 1500 caaggctatt tgaattagat gaaactcaag gcagcacaaa cagaattgtt gggacctatg 1560 gatatatggc accagaatat gcaatgcacg ggcaattttc tgtgaagtca gatgttttta 1620 gctttggagt actagtctta gaaattttaa gtggccaaaa aaacacttgt ttcagaaatg 1680 gagaatcggt ggaagacctt ctgagttttg cttggtcgag ctggcgtaat ggaacaacta 1740 taaattttgt agatccaatg ctgaaggaaa gcacaggact gattcgtgac ataatgagaa 1800 acattcacat agctttattg tgtgttcaag aaagtgtggc tgatagacca accatggcag 1860 ctgttgttct catgctcagt agcttttcgt tgagtcttcc catgccttca gggccagcat 1920 tctatatgca cagtaatatt accgcaggga cgtcgcttat tcaagaatac aacacaagag 1980 tgacagactc tagtgaacga gccaaaagta aatctattgg ttcatcacga aatgaggcgt 2040 ccataactga gttatatcct cgttaacatc tttagtataa tagccttttt ctccgatttt 2100 acttatttta cctgccttgg acttgtagtc tagctagcaa ttattgtaaa gttttattta 2160 acttgagcta aatacatcta tatccatttt aactggcttg ctttgtaaga tagcatcatt 2220 tcgccaaaaa aaaaaaaaaa aa 2242 7 2144 DNA Solanum tuberosum PRK-3 cDNA 7 atcaaaactt gcttgtaact taaacagaca tagccatggc tattcaaaag tggctactca 60 ttctgttttt gcatatacat gttctcaata ttgtagcgca gctgcctgat ttacgatttg 120 gttcatgtgg tggcaatggt aactatactg agaatagtac ctacaagaac aatctaaaca 180 cactcctcac ttccctttcc tcaaaaatag acaactatgg tttctacaat gcttctattg 240 gtcaaaactc tgatagagcc agcgttattg tgttatgtag aggagatgtg gagctagccg 300 attgtcgcgg ttgtgtcgat aatgttgttc aaaagattgc acagttatgt cctaaccaaa 360 aagaagtttt cggtggctat gatggatcga atgccacaaa acccgaggaa tttaaccagg 420 agctagggag attattggaa aacttacgag atcgtgctgt agatgatggt cctcttcaaa 480 aatatgctag tggtaatgcg acaggtccag attttcaggc tatatatgca cttgtgcagt 540 gtactcctga tttatctcgt cagagttgct tcagttgctt aagtgacgct tatggaaaca 600 tgcctcgctg tccctgcctg ggtaagaggg gtggaagaat cataggggtt aggtgcaact 660 tccgttatga aagttcccgt ttcttcgagg atgtgccgtt ggaagctcca ccacctgcag 720 gaaatgataa tacaacagtt ccaacaggga cggacgataa aacagttcca acaggagagg 780 atgacaaaac aacacgaaca atcatcatta tcgttgtgtc aactgttaca gttgttattc 840 ttattatttg tattgctgtc atcttgataa ggaggcgaaa gaggaagctg gtgaacgaaa 900 ttcagagtac atctgtagat gatactagta ttgcagaatc ttttcaatat gatttttcgg 960 caattagagc agcaacagat gacttctcag atgctaataa gctcggagaa ggcggatttg 1020 gtcctgtgta caagggtaag cttcaaaatg gacaagaagt agcagtgaaa aggttatcag 1080 cagattcagg ccaaggtgat ctagaattca aaaatgaggt cttgttggtt gccaggcttc 1140 aacacaggaa tttggttagg ttgctgggat tttgcctaga cggaacagag cgacttcttg 1200 tctatgagtt tgttcccaat gcaagtcttg accacttctt atttgattca gttaaacgta 1260 ggcaattgga ttgggaaagg cgatccaaaa tcataggagg cattgctaag ggaattcttt 1320 atcttcatga ggattctagg cttcggatca ttcaccgtga tctcaaagct agtaatgttc 1380 tactagatgc agaaatgaat cctaaaatct cagattttgg catggcaagg ctatttgaat 1440 tagatgaaac tcaaggcagc acaaacagaa ttgttgggac ctatggatat atggcaccag 1500 aatatgcaat gcacgggcaa ttttctgtga agtcagatgt ttttagcttt ggagtactag 1560 tcttagaaat tttaagtggc caaaaaaaca cttgtttcag aaatggagaa tcggtggaag 1620 accttctgag ttttgcttgg tcgagctggc gtaatggaac aactataaat tttgtagatc 1680 caatgctgaa ggaaagcaca ggactgattc gtgacataat gagaaacatt cacatagctt 1740 tattgtgtgt tcaagaaagt gtggctgata gaccaaccat ggcagctgtt gttctcatgc 1800 tcagtagctt ttcgttgagt cttcccatgc cttcagggcc agcattctat atgcacagta 1860 atattaccgc agggacgtcg cttattcaag aatacaacac aagagtgaca gactctagtg 1920 aacgagccaa aagtaaatct attggttcat cacgaaatga ggcgtccata actgagttat 1980 atcctcgtta acatctttag tataatagcc tttttctccg attttactta ttttacctgc 2040 cttggacttg tagtctagct agcaattatt gtaaagtttt atttaacttg agctaaatac 2100 atctatatcc attccaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 2144 8 2411 DNA Solanum tuberosum PRK-4 cDNA 8 aaaacttgct tgtaacttaa acagacatag ccatggctat tcaaaagtgg ctactcattc 60 tgtttttgca tatacatgtt ctcaatattg tagcgcagct gcctgattta cgatttggtt 120 catgtggtgg caatggtaac tatactgaga atagtaccta caagaacaat ctaaacacac 180 tcctcacttc cctttcctca aaaatagaca actatggttt ctacaatgct tctattggtc 240 aaaactctga tagagccagc gttattgtgt tatgtagagg agatgtggag ctagccgatt 300 gtcgcggttg tgtcgataat gttgttcaaa agattgcaca gttatgtcct aaccaaaaag 360 aagttttcgg tggctatgat ggatgtatgt tacagtattc aaatcaatcc attttagaaa 420 ctacatcatt ttctcttaaa tattactttt ggaatccagc gaatgccaca aaacccgagg 480 aatttaacca ggagctaggg agattattgg aaaacttacg agatcgtgct gtagatgatg 540 gtcctcttca aaaatatgct agtggtaatg cgacaggtcc agattttcag gctatatatg 600 cacttgtgca gtgtactcct gatttatctc gtcagagttg cttcagttgc ttaagtgacg 660 cttatggaaa catgcctcgc cgtccctgcc tgggtaagag gggtggaaga atcatagggg 720 ttaggtgcaa cttccgttat gaaagttccc gtttcttcga ggatgtgccg ttggaagctc 780 caccacctgc aggaaatgat aatacaacag ttccaacagg gacggacgat aaaacagttc 840 caacaggaga ggatgacaaa acaacacgaa caatcatcat tatcgttgtg tcaactgtta 900 cagttgttat tcttattatt tgtattgctg tcatcttgat aaggaggcga aagaggaagc 960 tggtgaacga aattcagagt acatctgtag atgatactag tattgcagaa tcttttcaat 1020 atgatttttc ggcaattaga gcagcaacag atgacttctc agatgctaat aagctcggag 1080 aaggcggatt tggtcctgtg tacaagggta agcttcaaaa tggacaagaa gtagcagtga 1140 aaaggttatc agcagattca ggccaaggtg atctagaatt caaaaatgag gtcttgttgg 1200 ttgccaggct tcaacacagg aatttggtta ggttgctggg attttgccta gacggaacag 1260 agcgacttct tgtctatgag tttgttccca atgcaagtct tgaccacttc ttatttgatt 1320 cggttaaacg taggcaattg gattgggaaa ggcgatccaa aatcatagga ggcattgcta 1380 agggaattct ttatcttcat gaggattcta ggcttcggat cattcaccgt gatctcaaag 1440 ctagtaatgt tctactagat gcagaaatga atcctaaaat ctcagatttt ggcatggcaa 1500 ggctatttga attagatgaa actcaaggca gcacaaacag aattgttggg acctatggat 1560 atatggcacc agaatatgca atgcacgggc aattttctgt gaagtcagat gtttttagct 1620 ttggagtact agtcttagaa attttaagtg gccaaaaaaa cacttgtttc agaaatggag 1680 aatcggtgga agaccttctg agttttgctt ggtcgagctg gcgtaatgga acaactataa 1740 attttgtaga tccaatgctg aaggaaagca caggactgat tcgtgacata atgagaaaca 1800 ttcacatagc tttattgtgt gttcaagaaa gtgtggctga tagaccaacc atggcagctg 1860 ttgttctcat gctcagtagc ttttcgttga gtcttcccat gccttcaggg ccagcattct 1920 atatgcacag taatattacc gcagggacgt cgcttattca agaatacaac acaagagtga 1980 cagactctag tgaacgagtc aaaagtaaat ctattggttc atcacgaaat gaggcgtcca 2040 taactgagtt atatcctcgt taacatcttt agaataatag cctttttctc cgattttact 2100 tattttacct gccttggact tgtagtctag ctagcaatta ttgtaaagtt ttatttaact 2160 tgagctaaat acatctatat ccattttaac tggcttgctt tgtaagatag catcatttcg 2220 gcatctcaca cgtcttttca actttatgga agaggcaaaa gggaaaagaa aacaagagga 2280 aaggaaaagg tctcaatttc tagaactcct tccgtacccc tctttcttta tgttattgaa 2340 gagccaaata acttttcgta ttctaaattc atataaagtt ttactccaaa aaaaaaaaaa 2400 aaaaaaaaaa a 2411 9 1975 DNA Arabidopsis thaliana Atpr3mia 9 atgtcttatt actcctcttt cttcttcctt ttcctcttct cctttctcac gagtttcaga 60 gtttctgctc aagatcccac ttacgtatac cacacctgtc aaaatacggc aaattacaca 120 agtaacagca cttacaacaa caatctcaag acccttttgg cttctctttc ttcccgcaac 180 gcttcccact ccaccggatt ccaaaacgcc accgtcggcc aagctccaga cagagtcacc 240 ggacttttca actgccgtgg agatgtctca acggaagttt gccgtagatg cgtctccttt 300 gccgtcaacg acaccttaac tcggtgtcct aaccagaaag aagccacact ttattacgat 360 gagtgtgtac ttagatactc taaccagaat attctctcga cactaataac caccggtgga 420 gttatcttgg ttaacacccg gaatgtgaca tctaaccaac tagacctgtt aagtgatttg 480 gttttgccta ctctgaacca agccgccacc gtagccttga acagttccaa aaattcggta 540 cgagaaaaaa caattttact gcgttgcaga gtttctatgg actggttcag tgcacacctg 600 atctcacgag acaagactgt tcgcgttgtc tccaactggt catcaatcaa atacctactg 660 acagaattgg tgcaagaatt attaatccga gctgtacttc aagatacgag atctacgcgt 720 tttacaccga atccgccgtt ccaccaccac caccaccacc gtcaatttct actcctccag 780 tttcagctcc tccgcgatct ggaaaagatg ggaattcaaa ggtgttagtg atagccattg 840 ttgtgcctat tatagtggct gttctgcttt ttatagctgg ttattgtttc cttacaagaa 900 gggcaaggaa gagttattac acaccatctg catttgctgg agatgatatc acaacagcag 960 actcactaca gcttgattat agaacaattc aaactgcaac agatgatttt gtagagagta 1020 ataagattgg tcaaggtgga tttggtgagg tttacaaggg tactttatcg gatgggactg 1080 aagttgcggt gaagcgacta tcaaagtcat caggacaagg tgaagtagag ttcaagaacg 1140 aggttgttct tgttgcaaag ctacaacata gaaatctggt tagacttctt gggttttgtc 1200 tagacggaga agaaagagta ctggtctacg agtatgtgcc caacaaaagt cttgattact 1260 tcctctttga ccctgcaaag aaaggccagc tggattggac tcgacgatac aagattatcg 1320 gtggagttgc tcgagggatt ctgtatcttc atcaagactc acgactcaca atcatacacc 1380 gtgacctcaa agccagtaac attctcttgg acgcggatat gaatcctaaa attgcagatt 1440 ttggaatggc taggatcttt ggattggacc aaaccgaaga gaacacaagc agaatagttg 1500 gtacctatgg ttacatgtct cccgagtatg caatgcacgg tcagtactca atgaaatctg 1560 atgtctatag cttcggagtg ttagttcttg agattataag tggcaagaag aatagcagct 1620 tctaccaaac agatggtgca catgacttgg tctcatatgc ttgggggctt tggagtaacg 1680 ggcgaccatt agaactcgtg gatccagcta ttgtagaaaa ttgccaaagg aatgaagttg 1740 ttcgatgtgt ccatatcggt cttttatgtg ttcaagaaga tcctgctgag cgtccgacgt 1800 tgtcaaccat tgttctgatg ctcactagta acactgtgac tttacccgtg ccatggcaac 1860 ccggtctttt ctttcagagt agaattggaa aagacccgct tgatacagat acaacgagca 1920 aatctcttct agggtctgtt gacgatgcat ccatcacaga tatacatcct cgatg 1975 10 278 PRT Solanum tuberosum 10 Met Ala Ile Gln Lys Trp Leu Leu Phe Leu Phe Leu His Leu His Val 1 5 10 15 Leu Asn Ile Val Ala Gln Leu Pro Asp Leu Arg Phe Gly Ile Cys Gly 20 25 30 Lys Ser Gly Asn Tyr Thr Glu Asn Ser Thr Tyr Lys Asn Asp Leu Asn 35 40 45 Thr Leu Leu Thr Ser Leu Ser Ser Lys Ile Asp Lys Tyr Gly Phe Tyr 50 55 60 Asn Ala Ser Ile Gly Gln Asn Ser Asp Arg Ala Ser Val Ile Val Leu 65 70 75 80 Cys Arg Gly Asp Val Glu Leu Asp Asp Cys Arg Gly Cys Val Asp Asn 85 90 95 Val Val Gln Lys Ile Ala Gln Leu Cys Pro Asn Gln Lys Glu Val Phe 100 105 110 Gly Gly Tyr Asp Gly Cys Met Leu Gln Tyr Ser Asn Gln Ser Ile Ile 115 120 125 Asp Thr Pro Ser Leu Ser Val Gln Leu Phe Leu Trp Asn Thr Ala Asn 130 135 140 Ala Ser Lys Pro Glu Glu Phe Asn Gln Glu Leu Gly Lys Leu Leu Glu 145 150 155 160 Asn Leu Arg Asp Arg Ala Ala Gln Gly Gly Pro Leu Gln Lys Tyr Ala 165 170 175 Thr Gly Thr Thr Ile Gly Pro Asp Ile Gln Pro Ile Tyr Ala Leu Val 180 185 190 Gln Cys Thr Pro Asp Leu Ser Arg Gln Ser Cys Phe Asp Cys Leu Thr 195 200 205 Asp Ala Tyr Gly Thr Leu Pro Gln Cys Pro Cys Leu Gly Lys Thr Gly 210 215 220 Gly Arg Ile Ile Gly Ile Arg Cys Asn Phe Arg Tyr Glu Ile Ser Arg 225 230 235 240 Phe Phe Val Asp Val Pro Leu Glu Ala Pro Pro Pro Ala Gly Asn Asp 245 250 255 Asn Lys Thr Val Pro Thr Gly Thr Glu Asn Lys Thr Pro Pro Thr Gly 260 265 270 Lys Asp Asp Lys Thr Thr 275 11 278 PRT Solanum tuberosum 11 Met Ala Ile Gln Lys Trp Leu Leu Ile Leu Phe Leu His Ile His Val 1 5 10 15 Leu Asn Ile Val Ala Gln Leu Pro Asp Leu Arg Phe Gly Ser Cys Gly 20 25 30 Gly Asn Gly Asn Tyr Thr Glu Asn Ser Thr Tyr Lys Asn Asn Leu Asn 35 40 45 Thr Leu Leu Thr Ser Leu Ser Ser Lys Ile Asp Asn Tyr Gly Phe Tyr 50 55 60 Asn Ala Ser Ile Gly Gln Asn Ser Asp Arg Ala Ser Val Ile Val Leu 65 70 75 80 Cys Arg Gly Asp Val Glu Leu Ala Asp Cys Arg Gly Cys Val Asp Asn 85 90 95 Val Val Gln Lys Ile Ala Gln Leu Cys Pro Asn Gln Lys Glu Val Phe 100 105 110 Gly Gly Tyr Asp Gly Cys Met Leu Gln Tyr Ser Asn Gln Ser Ile Leu 115 120 125 Glu Thr Thr Ser Phe Ser Leu Lys Tyr Tyr Phe Trp Asn Pro Ala Asn 130 135 140 Ala Thr Lys Pro Gly Glu Phe Asn Gln Glu Leu Gly Arg Leu Leu Glu 145 150 155 160 Asn Leu Arg Asp Arg Ala Val Asp Asp Gly Pro Leu Gln Lys Tyr Ala 165 170 175 Ser Gly Asn Ala Thr Gly Pro Asp Phe Gln Ala Ile Tyr Ala Leu Val 180 185 190 Gln Cys Thr Pro Asp Leu Ser Arg Gln Ser Cys Phe Ser Cys Leu Ser 195 200 205 Asp Ala Tyr Gly Asn Met Pro Arg Cys Pro Cys Leu Gly Lys Arg Gly 210 215 220 Gly Arg Ile Ile Gly Val Arg Cys Asn Phe Arg Tyr Glu Ser Ser Arg 225 230 235 240 Phe Phe Glu Asp Val Pro Leu Glu Ala Pro Pro Pro Ala Gly Asn Asp 245 250 255 Asn Thr Thr Val Pro Thr Gly Thr Asp Asp Lys Thr Val Pro Thr Gly 260 265 270 Glu Asp Asp Lys Thr Thr 275 12 253 PRT Solanum tuberosum 12 Met Ala Ile Gln Lys Trp Leu Leu Ile Leu Phe Leu His Ile His Val 1 5 10 15 Leu Asn Ile Val Ala Gln Leu Pro Asp Leu Arg Phe Gly Ser Cys Gly 20 25 30 Gly Asn Gly Asn Tyr Thr Glu Asn Ser Thr Tyr Lys Asn Asn Leu Asn 35 40 45 Thr Leu Leu Thr Ser Leu Ser Ser Lys Ile Asp Asn Tyr Gly Phe Tyr 50 55 60 Asn Ala Ser Ile Gly Gln Asn Ser Asp Arg Ala Ser Val Ile Val Leu 65 70 75 80 Cys Arg Gly Asp Val Glu Leu Ala Asp Cys Arg Gly Cys Val Asp Asn 85 90 95 Val Val Gln Lys Ile Ala Gln Leu Cys Pro Asn Gln Lys Glu Val Phe 100 105 110 Gly Gly Tyr Asp Gly Ser Asn Ala Thr Lys Pro Glu Glu Phe Asn Gln 115 120 125 Glu Leu Gly Arg Leu Leu Glu Asn Leu Arg Asp Arg Ala Val Asp Asp 130 135 140 Gly Pro Leu Gln Lys Tyr Ala Ser Gly Asn Ala Thr Gly Pro Asp Phe 145 150 155 160 Gln Ala Ile Tyr Ala Leu Val Gln Cys Thr Pro Asp Leu Ser Arg Gln 165 170 175 Ser Cys Phe Ser Cys Leu Ser Asp Ala Tyr Gly Asn Met Pro Arg Cys 180 185 190 Pro Cys Leu Gly Lys Arg Gly Gly Arg Ile Ile Gly Val Arg Cys Asn 195 200 205 Phe Arg Tyr Glu Ser Ser Arg Phe Phe Glu Asp Val Pro Leu Glu Ala 210 215 220 Pro Pro Pro Ala Gly Asn Asp Asn Thr Thr Val Pro Thr Gly Thr Asp 225 230 235 240 Asp Lys Thr Val Pro Thr Gly Glu Asp Asp Lys Thr Thr 245 250 13 278 PRT Solanum tuberosum 13 Met Ala Ile Gln Lys Trp Leu Leu Ile Leu Phe Leu His Ile His Val 1 5 10 15 Leu Asn Ile Val Ala Gln Leu Pro Asp Leu Arg Phe Gly Ser Cys Gly 20 25 30 Gly Asn Gly Asn Tyr Thr Glu Asn Ser Thr Tyr Lys Asn Asn Leu Asn 35 40 45 Thr Leu Leu Thr Ser Leu Ser Ser Lys Ile Asp Asn Tyr Gly Phe Tyr 50 55 60 Asn Ala Ser Ile Gly Gln Asn Ser Asp Arg Ala Ser Val Ile Val Leu 65 70 75 80 Cys Arg Gly Asp Val Glu Leu Ala Asp Cys Arg Gly Cys Val Asp Asn 85 90 95 Val Val Gln Lys Ile Ala Gln Leu Cys Pro Asn Gln Lys Glu Val Phe 100 105 110 Gly Gly Tyr Asp Gly Cys Met Leu Gln Tyr Ser Asn Gln Ser Ile Leu 115 120 125 Glu Thr Thr Ser Phe Ser Leu Lys Tyr Tyr Phe Trp Asn Pro Ala Asn 130 135 140 Ala Thr Lys Pro Glu Glu Phe Asn Gln Glu Leu Gly Arg Leu Leu Glu 145 150 155 160 Asn Leu Arg Asp Arg Ala Val Asp Asp Gly Pro Leu Gln Lys Tyr Ala 165 170 175 Ser Gly Asn Ala Thr Gly Pro Asp Phe Gln Ala Ile Tyr Ala Leu Val 180 185 190 Gln Cys Thr Pro Asp Leu Ser Arg Gln Ser Cys Phe Ser Cys Leu Ser 195 200 205 Asp Ala Tyr Gly Asn Met Pro Arg Arg Pro Cys Leu Gly Lys Arg Gly 210 215 220 Gly Arg Ile Ile Gly Val Arg Cys Asn Phe Arg Tyr Glu Ser Ser Arg 225 230 235 240 Phe Phe Glu Asp Val Pro Leu Glu Ala Pro Pro Pro Ala Gly Asn Asp 245 250 255 Asn Thr Thr Val Pro Thr Gly Thr Asp Asp Lys Thr Val Pro Thr Gly 260 265 270 Glu Asp Asp Lys Thr Thr 275 14 658 PRT Arabidopsis thaliana 14 Met Ser Tyr Tyr Ser Ser Phe Phe Phe Leu Phe Leu Phe Ser Phe Leu 1 5 10 15 Thr Ser Phe Arg Val Ser Ala Gln Asp Pro Thr Tyr Val Tyr His Thr 20 25 30 Cys Gln Asn Thr Ala Asn Tyr Thr Ser Asn Ser Thr Tyr Asn Asn Asn 35 40 45 Leu Lys Thr Leu Leu Ala Ser Leu Ser Ser Arg Asn Ala Ser His Ser 50 55 60 Thr Gly Phe Gln Asn Ala Thr Val Gly Gln Ala Pro Asp Arg Val Thr 65 70 75 80 Gly Leu Phe Asn Cys Arg Gly Asp Val Ser Thr Glu Val Cys Arg Arg 85 90 95 Cys Val Ser Phe Ala Val Asn Asp Thr Leu Thr Arg Cys Pro Asn Gln 100 105 110 Lys Glu Ala Thr Leu Tyr Tyr Asp Glu Cys Val Leu Arg Tyr Ser Asn 115 120 125 Gln Asn Ile Leu Ser Thr Leu Ile Thr Thr Gly Gly Val Ile Leu Val 130 135 140 Asn Thr Arg Asn Val Thr Ser Asn Gln Leu Asp Leu Leu Ser Asp Leu 145 150 155 160 Val Leu Pro Thr Leu Asn Gln Ala Ala Thr Val Ala Leu Asn Ser Ser 165 170 175 Lys Lys Phe Gly Thr Arg Lys Asn Asn Phe Thr Ala Leu Gln Ser Phe 180 185 190 Tyr Gly Leu Val Gln Cys Thr Pro Asp Leu Thr Arg Gln Asp Cys Ser 195 200 205 Arg Cys Leu Gln Leu Val Ile Asn Gln Ile Pro Thr Asp Arg Ile Gly 210 215 220 Ala Arg Ile Ile Asn Pro Ser Cys Thr Ser Arg Tyr Glu Ile Tyr Ala 225 230 235 240 Phe Tyr Thr Glu Ser Ala Val Pro Pro Pro Pro Pro Pro Pro Ser Ile 245 250 255 Ser Thr Pro Pro Val Ser Ala Pro Pro Arg Ser Gly Lys Asp Gly Asn 260 265 270 Ser Lys Val Leu Val Ile Ala Ile Val Val Pro Ile Ile Val Ala Val 275 280 285 Leu Leu Phe Ile Ala Gly Tyr Cys Phe Leu Thr Arg Arg Ala Arg Lys 290 295 300 Ser Tyr Tyr Thr Pro Ser Ala Phe Ala Gly Asp Asp Ile Thr Thr Ala 305 310 315 320 Asp Ser Leu Gln Leu Asp Tyr Arg Thr Ile Gln Thr Ala Thr Asp Asp 325 330 335 Phe Val Glu Ser Asn Lys Ile Gly Gln Gly Gly Phe Gly Glu Val Tyr 340 345 350 Lys Gly Thr Leu Ser Asp Gly Thr Glu Val Ala Val Lys Arg Leu Ser 355 360 365 Lys Ser Ser Gly Gln Gly Glu Val Glu Phe Lys Asn Glu Val Val Leu 370 375 380 Val Ala Lys Leu Gln His Arg Asn Leu Val Arg Leu Leu Gly Phe Cys 385 390 395 400 Leu Asp Gly Glu Glu Arg Val Leu Val Tyr Glu Tyr Val Pro Asn Lys 405 410 415 Ser Leu Asp Tyr Phe Leu Phe Asp Pro Ala Lys Lys Gly Gln Leu Asp 420 425 430 Trp Thr Arg Arg Tyr Lys Ile Ile Gly Gly Val Ala Arg Gly Ile Leu 435 440 445 Tyr Leu His Gln Asp Ser Arg Leu Thr Ile Ile His Arg Asp Leu Lys 450 455 460 Ala Ser Asn Ile Leu Leu Asp Ala Asp Met Asn Pro Lys Ile Ala Asp 465 470 475 480 Phe Gly Met Ala Arg Ile Phe Gly Leu Asp Gln Thr Glu Glu Asn Thr 485 490 495 Ser Arg Ile Val Gly Thr Tyr Gly Tyr Met Ser Pro Glu Tyr Ala Met 500 505 510 His Gly Gln Tyr Ser Met Lys Ser Asp Val Tyr Ser Phe Gly Val Leu 515 520 525 Val Leu Glu Ile Ile Ser Gly Lys Lys Asn Ser Ser Phe Tyr Gln Thr 530 535 540 Asp Gly Ala His Asp Leu Val Ser Tyr Ala Trp Gly Leu Trp Ser Asn 545 550 555 560 Gly Arg Pro Leu Glu Leu Val Asp Pro Ala Ile Val Glu Asn Cys Gln 565 570 575 Arg Asn Glu Val Val Arg Cys Val His Ile Gly Leu Leu Cys Val Gln 580 585 590 Glu Asp Pro Ala Glu Arg Pro Thr Leu Ser Thr Ile Val Leu Met Leu 595 600 605 Thr Ser Asn Thr Val Thr Leu Pro Val Pro Trp Gln Pro Gly Leu Phe 610 615 620 Phe Gln Ser Arg Ile Gly Lys Asp Pro Leu Asp Thr Asp Thr Thr Ser 625 630 635 640 Lys Ser Leu Leu Gly Ser Val Asp Asp Ala Ser Ile Thr Asp Ile His 645 650 655 Pro Arg 

What is claimed is:
 1. Isolated nucleic acid sequences comprising polynucleotides encoding receptor-like protein kinases, which comprise preferably in the extracellular domain one or more cystein repeats, characterized in that potato receptor-like kinase (PRK)-like nucleic acid sequences are capable of encoding potato receptor-like kinases (PRKs) substantially homologous to gene products of PRK (SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4) or fragments or derivatives thereof and having substantially the same properties or functions as SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4.
 2. The nucleic acid sequences according to claim 1, characterized in that the potato receptor-like kinase (PRK)-like nucleic acid sequences are capable of hybridizing with SEQ ID:5, SEQ ID:6, SEQ ID:7 or SEQ ID:8 or complementary strands thereof under defined conditions.
 3. The nucleic acid sequences according to claim 1, characterized in that they comprise SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4 obtainable from potato under defined conditions.
 4. The nucleic acid sequences according to claim 1, characterized in that the extracellular domains comprise a conserved bi-modular pattern of one or more cysteine repeats.
 5. The nucleic acid sequences according to claim 1, characterized in that the expression of genes encoding for gene products SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4 is induced by Erwinia carotovora.
 6. The nucleic acid sequences according to claim 1, characterized in that the expression of genes encoding for gene products SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4 is induced by oligouronides.
 7. The nucleic acid sequences according to claim 1, characterized in that potato receptor-like kinases (PRKs) function as receptors for ligands released during plant stress conditions by pathogens.
 8. The nucleic acid sequences according to claim 1, characterized in that potato receptor-like kinases (PRKs) are formed by alternative splicing.
 9. The nucleic acid sequences according to claim 1, characterized in that potato receptor-like kinases (PRKs) are involved in signal perception during potato defense responses against Erwinia carotovora.
 10. The nucleic acid sequences according to claim 1, characterized in that the expression of potato receptor-like kinases (PRKs) is incuced by response to elicitors.
 11. The nucleic acid sequences according to claim 10, characterized in that the elicitors are oligouronides.
 12. The nucleic acid sequences according to claim 10, characterized in that the elicitors are oligogalacturonides.
 13. Potato receptor-like kinase (PRK)-like gene products expressed by the nucleic acid sequences according to claims 1, characterized in that the PRK-like gene products are polypeptides comprising in their extracellular domains a conserved bi-modular patterns of one or more cysteine repeats.
 14. The potato receptor-like kinase (PRK)-like gene products according to claim 13, characterized in that they comprise polypeptides having amino acid sequences substantially homologous with SEQ ID:1, SEQ ID:2, SEQ ID:3 or SEQ ID:4.
 15. The potato receptor-like kinase (PRK)-like gene products according to claim 13, characterized in that PRK-like gene products are polypeptides substantially similar to the gene products encoded by SEQ ID:5, SEQ ID:6, SEQ ID:7 or SEQ ID:8.
 16. A method for conferring resistance to pathogens in a plant, characterized in that the method comprises introducing into the plant a recombinant expression construct comprising a plant promoter operably linked to a potato receptor-like kinase (PRK)-like nucleotide sequence of claim 1 or derivatives or fragments thereof.
 17. A method for conferring resistance to pathogens in a plant, characterized in that the method comprises introducing into the plant a recombinant expression construct comprising a plant promoter operably linked to a Arabidopsis receptor-like kinase (PRK)-like nucleotide sequence SEQ ID NO: 9 or derivatives or fragments thereof.
 18. The method of claim 16, characterized in that potato receptor-like kinase (PRK)-like nucleotide sequence encodes a potato receptor-like kinase (PRK)-like gene product of claim
 14. 19. The method of claim 17, characterized in that Arabidopsis receptor-like kinase (PRK)-like nucleotide sequence encodes a Arabidopsis receptor-like kinase (PRK)-like gene product of SEQ ID NO:14.
 20. The method of claim 16, characterized in that the plant tissue is from potato.
 21. The method of claim 16, characterized in that the plant tissue is from Arabidopsis.
 22. A DNA construct for cloning and/or transforming plants, wherein the DNA construct comprises the DNA sequences of claim 1 functionally combined with regulatory sequences.
 23. An expression vector comprising the nucleotide sequence of claim
 1. 24. A host cell comprising the DNA construct of claim
 20. 